Information on EC 3.1.27.3 - ribonuclease T1

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
3.1.27.3
-
RECOMMENDED NAME
GeneOntology No.
ribonuclease T1
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
mechanism
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
hydrolysis of the phosphodiester bonds of single-stranded RNA at the 3'-side of guanosine nucleotides occurs with high specificity
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
the conjugate base of the gamma-carboxyl group of Clu46 is required for tight binding of guanine and hypoxanthine moieties at the primary recognition site of the enzyme
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
small single-domain protein of 104 amino acid residues that cleaves single-stranded RNA specifically at the 3'-side of guanine residues in a two-step mechanism
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
the enzyme is composed of 104 amino acid residues and it cleaves single-stranded RNA specifically after guanosine residues to produce mono or oligo-ribonucleotides with terminal 3'-phosphate
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
the reaction is a transphosphorylation yielding a 2',3'-cyclophosphate. This cyclic intermediate is hydrolyzed by a water molecule to a 3'-phosphate in a separate step which is the reversal of the transphosphorylation. The transphosphorylation consists of a nucleophilic in-line inversion displacement at the phosphorous atom of the 5'-leaving group by the entering 2'-oxygen. Tyr38, His40, Glu58, His92 and Phe100 are involved in the catalysis of RNA degradation. During cyclization, Glu58 and His92 serve as the catalytic base and acid, respectively
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
the side chain carboxyl of Glu54 accepts a proton from the ribose 2'OH group and the protonated imidazole ring of His85 donates a proton to the leaving 5'O group. The positively charged side chain of Arg65 is to promote the formation of a negatively charged, pentacovalent intermediate state of the phosphate group
-
two-stage endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in Gp with 2',3'-cyclic phosphate intermediates
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
-
-
hydrolysis of phosphoric ester
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Aspergillus oryzae ribonuclease
-
-
-
-
Binase
-
-
-
-
Binase
P00649
-
EC 2.7.7.26
-
-
formerly
-
EC 3.1.4.8
-
-
formerly
-
Gln25-RNase T1
-
two isoforms of RNase T1 exist in nature: one has Gln and the other Lys at position 25
guanyl-preferring RNase
-
-
Guanyl-specific RNase
-
-
-
-
guanylate endoribonuclease
-
-
Nuclease, guanyloribo-
-
-
-
-
Nuclease, ribo-, Aspergillus oryzae
-
-
-
-
ribonuclease A
-
-
Ribonuclease C2
-
-
-
-
Ribonuclease Ch
-
-
-
-
ribonuclease E/G
-
-
Ribonuclease F1
-
-
-
-
ribonuclease G
-
-
Ribonuclease guaninenucleotido-2'-transferase (cyclizing)
-
-
-
-
Ribonuclease N1
-
-
-
-
Ribonuclease N3
-
-
-
-
ribonuclease NT
-
-
ribonuclease Pb2
-
-
Ribonuclease PP1
-
-
-
-
Ribonuclease SA
-
-
-
-
ribonuclease Sa2
-
-
ribonuclease T1
P00651
-
Ribonuclease U1
-
-
-
-
RNase A
-
-
RNase F1
-
-
-
-
RNase Fl1
-
-
-
-
RNase Fl2
-
-
-
-
RNase G
-
-
-
-
RNase G
Escherichia coli N3433
-
-
-
RNase Ms
-
-
-
-
RNase N1
-
-
-
-
RNase N1
P09646
-
RNase N2
-
-
-
-
RNase Pb1
-
-
-
-
RNase Pc
-
-
-
-
RNase Po1
-
-
-
-
RNase Sa
-
-
-
-
RNase Sa3
-
-
-
-
RNase St
-
-
-
-
RNase T1
-
-
-
-
RNase T1
P00651
-
RNase T1 RV
-
mutant RNase T1 K41E/Y42F/N43R/Y45W/E46N
RNase T1-R2
-
mutant RNase T1 K41E/Y42F/N43R/Y45W/E46N/W59Y
RNase Th1
-
-
-
-
RNase U1
-
-
-
-
Guanyloribonuclease
-
-
-
-
additional information
-
the enzyme belongs to the superfamily of microbial N1/T1 RNases
CAS REGISTRY NUMBER
COMMENTARY
9026-12-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
similar enzyme
-
-
Manually annotated by BRENDA team
2 forms: T1-A, T1-B
-
-
Manually annotated by BRENDA team
strain 7P
SwissProt
Manually annotated by BRENDA team
Bacillus intermedius 7P
strain 7P
SwissProt
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
Escherichia coli N3433
gene rng
-
-
Manually annotated by BRENDA team
overview
-
-
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
deletion of gene rng leads to accumulation of 23S rRNA precursors in Escherichia coli
malfunction
Escherichia coli N3433
-
deletion of gene rng leads to accumulation of 23S rRNA precursors in Escherichia coli
-
metabolism
-
distinct roles of RNase E and RNase G in mRNA decay and tRNA processing, overview
physiological function
-
binase is a regulator of RNA-dependent processes of cell proliferation and apoptosis. Binase affects the total amount of intracellular RNA and the expression of proapoptotic and antiapoptotic mRNAs
physiological function
-
RNase G is involved in the maturation of the 5' terminus of 16S rRNA, the processing of a few tRNAs, and the initiation of decay of a limited number of mRNAs but is not required for cell viability and cannot substitute for RNase E under normal physiological conditions
physiological function
-
barnase can help the population to win the competition with other bacteria for ecological niches, acting as a toxin. Toxic extracellular RNases and antitoxic barstar build an analogous system
physiological function
-
binase can help the population to win the competition with other bacteria for ecological niches, acting as a toxin
physiological function
-
RNase G is involved in maturation of the 5' end of the 23S rRNA processing the 5' region by cleaving the 77 extra nucleotides at the 5' end
physiological function
Escherichia coli N3433
-
RNase G is involved in maturation of the 5' end of the 23S rRNA processing the 5' region by cleaving the 77 extra nucleotides at the 5' end
-
metabolism
-
binase is involved in phosphate metabolism, low concentrations of extracellular inorganic phosphate induce the expression of phosphate regulon, Pho, genes, as well as the binase gene, binase expression is strongly dependent on a functional PhoP-PhoR two-component system
additional information
-
neither the native nor N-terminal extended form of RNase G can restore the growth defect associated with either the rne-1 or rneD1018 alleles, encoding RNase E, even when expressed at very high protein levels. In contrast, two distinct spontaneously derived single amino acid substitutions within the predicted RNase H domain of RNase G, generating the rng-219 and rng-248 alleles, result in complementation of the growth defect associated with various RNase E mutants. Complementation of the growth defect associated with RNase E-deficient strains is dependent on the intracellular level of the Rng-219 and Rng-248 proteins
additional information
-
comparison with binase from Bacillus pumilus, overview. Barnase expression is strictly dependent on activating Spo0A, the Spo0A protein is a multifunctional regulator that controls stress-related processes, such as sporulation, biofilm formation and cannibalism
additional information
-
comparison with barnase from Bacillus amyloliquefaciens, overview. Mutation in resD leads to a significant decrease in binase production, whereas mutation in spo0A causes its hyperproduction. Expression of binase is possible only in Spo0A-OFF cells
additional information
-
analysis of cumulative permeation and skin deposition of negatively charged RNAse T1 using porcine ear skin. RNAse T1 permeation is dependent upon current density,while skin deposition is not. RNAse T1 retains structural integrity and enzymatic function postiontophoresis. RNAse T1 appears to be bound to the epidermis alone
additional information
-
effects of water-water hydrogen bonding types upon the activity of the enzyme ribonuclease t1 through perturbation of the water hydrogen bonding distribution by using various salts, overview. Various salts differ in their ability to reduce the enzymatic activity of ribonuclease t1 correlated with the ability of each salt to promote high-angle hydrogen bonding in water. Increasing the population of high-angle hydrogen bonds among water molecules stabilizes the more compact, less active conformations of the enzyme
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
adenylyl-3',5'-cytidine
adenosine-2',3'-cyclic phosphate + cytidine
show the reaction diagram
-
ApC
-
-
?
adenylyl-3',5'-cytidine + H2O
adenosine 3'-phosphate + cytidine
show the reaction diagram
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, 195 for mutant enzyme E46N and 137 for mutant enzyme E46N/Y45W
-
-
?
adenylyl-3',5'-cytidine + H2O
adenosine 3'-phosphate + cytidine
show the reaction diagram
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, and 39 for mutant enzyme K41E/Y42F/N43R/Y45W/E46N/W59Y (RNase T1-R2) and mutant enzyme K41E/Y42F/N43R/Y45W/E46N (RNase RV)
-
-
?
ApC + H2O
adenosine 3'-phosphate + cytidine
show the reaction diagram
-
-
-
-
?
GpC + H2O
guanosine 3'-phosphate + cytidine
show the reaction diagram
-
-
-
-
?
GpC + H2O
?
show the reaction diagram
-
enzyme-catalyzed hydrolysis of GpC
-
-
?
GpU + H2O
guanosine 3'-phosphate + uridine
show the reaction diagram
-
-
-
-
?
GpU + H2O
guanosine 3'-phosphate + uridine
show the reaction diagram
-
hydrolysis
-
-
?
guanosine 2',3'-cyclic phosphate + H2O
guanosine-3'-phosphate
show the reaction diagram
-
-
-
-
-
guanosine 2',3'-cyclic phosphate + H2O
guanosine-3'-phosphate
show the reaction diagram
-
Ustilago sphaerogena, RNase U1
-
-
-
guanosine-2',3'-cyclic phosphate + H2O
guanosine 3'-phosphate
show the reaction diagram
-
hydrolysis
-
-
?
guanylyl(3',5') adenosine + H2O
?
show the reaction diagram
P00648
shift in nucleotide conformational equilibrium contributes to increased rate of catalysis of guanylyl(3',5') adenosine 3'-monophosphate versus guanylyl(3',5') adenosine
-
-
?
guanylyl(3',5') adenosine 3'-monophosphate + H2O
?
show the reaction diagram
P00648
shift in nucleotide conformational equilibrium contributes to increased rate of catalysis of guanylyl(3',5') adenosine 3'-monophosphate versus guanylyl(3',5') adenosine
-
-
?
guanylyl-3',5'-cytidine
guanosine-2',3'-cyclic phosphate + cytidine
show the reaction diagram
-
Transphosphorylation reaction
-
-
?
guanylyl-3',5'-cytidine + H2O
guanosine 3'-phosphate + cytidine
show the reaction diagram
-
GpC
-
-
?
guanylyl-3',5'-cytidine + H2O
?
show the reaction diagram
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, 195 for mutant enzyme E46N and 137 for mutant enzyme E46N/Y45W
-
-
?
guanylyl-3',5'-cytidine + H2O
?
show the reaction diagram
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, and 39 for mutant enzyme K41E/Y42F/N43R/Y45W/E46N/W59Y (RNase T1-R2) and mutant enzyme K41E/Y42F/N43R/Y45W/E46N (RNase RV)
-
-
?
guanylyl-3',5'-uridine
guanosine-2',3'-cyclic phosphate + uridine
show the reaction diagram
-
Transphosphorylation reaction with GpU or the diastereomers resulting from thio-substitution of a nonbridging oxygen of GpU, RpGp(S)U and SpGp(S)U as substrates. SpGp(S)U is a very poor substrate for wild type enzyme
-
-
?
poly(I) + H2O
?
show the reaction diagram
-
-
-
-
?
polyinosinic acid + H2O
?
show the reaction diagram
-
polyI
-
-
?
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
-
-
?
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
-
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
essential requirements: keto group at 6 position, trivalent nitrogen at 7 position of purine base
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
essential requirements: keto group at 6 position, trivalent nitrogen at 7 position of purine base
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
first step (r): cleavage of internucleotide bonds between 3'-guanylic acid groups and the 5'-hydroxyl groups of the adjacent nucleotides with the intermediary formation of guanosine 2',3'-cyclic phosphate, second step (ir): hydrolysis of cyclic phosphate to produce 3'-guanylate
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
first step (r): cleavage of internucleotide bonds between 3'-guanylic acid groups and the 5'-hydroxyl groups of the adjacent nucleotides with the intermediary formation of guanosine 2',3'-cyclic phosphate, second step (ir): hydrolysis of cyclic phosphate to produce 3'-guanylate
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
specificity
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
Hydrolyzed RNA
show the reaction diagram
-
specificity
3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp
-
RNA + H2O
?
show the reaction diagram
-
two-stage endonucleolytic cleavage to 3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp with 2,3'-cyclic phosphate intermediates
-
-
-
RNA + H2O
?
show the reaction diagram
-
identification of RNase T1 cleavage fragments of Bacillus megaterium ribosomal 5S RNA
-
-
?
guanylyl-3',5'-uridine + H2O
guanosine 3'-phosphate + uridine
show the reaction diagram
-
GpU
-
-
?
additional information
?
-
-
synthesis of guanylylnucleosides, oligoguanylate, guanosine-containing oligonucleotides with (3'-5')phosphodiester bonds, synthetic reaction can lead to the formation of unnatural (2'-5')-phosphodiester bonds, not: DNA, L-guanosine 2',3'-cyclic phosphate, L-inosine 2',3'-cyclic phosphate
-
-
-
additional information
?
-
-
heteroduplex of ribooligonucleotides are substrates for the enzyme
-
-
-
additional information
?
-
-
endonucleolytic activity is assayed using derivates of BR13, 5'-GGGACAGUAUUUG-3', a model oligonucleotide substrate for studies of RNase E and RNase G
-
-
-
additional information
?
-
-
the backbone dynamics of RNase T1, complexed with the productive substrate exo-cGPS isomer, in comparison to the RNase T1, complexed with the nonproductive substrate 3'GMP, is analyzed
-
-
-
additional information
?
-
-
in vivo binase affects the quantity of proapoptotic and antiapoptotic mRNAs, as expression of the p53 and hSK4 genes is increased and expression of the bcl-2 gene is reduced
-
-
-
additional information
?
-
-
barnase catalyses the overall hydrolysis of single-stranded RNA preferentially at guanylyl residues, yielding new guanosine 3'-phosphate and 5'-OH ends, in a two-step process with cleavage of the RNA chain by transesterification of a 5'-phosphoester bond to form a guanosine 2',3'-cyclic phosphate terminus in the first step, followed by its hydrolysis to a 3'-phosphate product in the second independent step
-
-
-
additional information
?
-
-
binase catalyses the overall hydrolysis of single-stranded RNA preferentially at guanylyl residues, yielding new guanosine 3'-phosphate and 5'-OH ends, in a two-step process with cleavage of the RNA chain by transesterification of a 5'-phosphoester bond to form a guanosine 2',3'-cyclic phosphate terminus in the first step, followed by its hydrolysis to a 3'-phosphate product in the second independent step
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
RNA + H2O
?
show the reaction diagram
-
two-stage endonucleolytic cleavage to 3'-phosphomononucleotides and 3'-phosphooligonucleotides ending in Gp with 2,3'-cyclic phosphate intermediates
-
-
-
additional information
?
-
-
barnase catalyses the overall hydrolysis of single-stranded RNA preferentially at guanylyl residues, yielding new guanosine 3'-phosphate and 5'-OH ends, in a two-step process with cleavage of the RNA chain by transesterification of a 5'-phosphoester bond to form a guanosine 2',3'-cyclic phosphate terminus in the first step, followed by its hydrolysis to a 3'-phosphate product in the second independent step
-
-
-
additional information
?
-
-
binase catalyses the overall hydrolysis of single-stranded RNA preferentially at guanylyl residues, yielding new guanosine 3'-phosphate and 5'-OH ends, in a two-step process with cleavage of the RNA chain by transesterification of a 5'-phosphoester bond to form a guanosine 2',3'-cyclic phosphate terminus in the first step, followed by its hydrolysis to a 3'-phosphate product in the second independent step
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
the enzyme has six binding sites for the cation
H+
-
the cation produces two different tautomers of the histidine residues His53 and His85
Sr2+
-
the enzyme contains a divalent ion-binding site involving Asp49, with preference for this cation. The ion binding at this site stabilizes the protein
additional information
-
Chalaropsis sp., no metal ion requirement
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
3'-N-hydroxyurea-3'-deoxythymidine-5'-phosphate
-
competitive, inhibition is enhanced by nearly 10fold in presence of Zn2 in wild-type enzyme, no enhancement in mutant enzyme E54Q
Acetic anhydride
-
-
barstar
-
dimerization can compete with binase inhibition by barstar, the ratio of competitve reaction rates depends on pH and ionic strength of the solution
-
barstar
P00649
-
-
barstar
-
a small protein of 89 amino acids, specifically inhibits barnase, barnase and barstar form a noncovalent one-to-one complex. Barstar inhibits barnase activity by sterically blocking its active site with an alpha-helix and adjacent loop. The association is stabilized by charge interaction involving the positively charged amino acid residues Lys27, Arg59, Arg83 and Arg87 of barnase and the negatively charged Asp35, Asp39 and Glu76 of barstar
-
Bromoacetic acid
-
-
Ca2+
-
Neurospora crassa RNase N4
Cu2+
-
Neurospora crassa RNase N4
Fe3+
-
Aspergillus niger RNase II
-
Guanylyl-2',5'-guanosine
-
-
Hg2+
-
Neurospora crassa RNase N4
iodoacetate
-
Neurospora crassa RNase N4
iodoacetate
-
Ribunuclease U1
Mg2+
-
Neurospora crassa RNase N4
Ozone
-
loss of activity at pH 7.5, at pH 4.75 the enzyme retains a decreased but distinct enzyme activity towards RNA without alteration of substrate specificity
Phenylglyoxal
-
-
Substrate analogs
-
-
-
Tosylglycolate
-
protection by substrate analogs
Zn2+
-
Neurospora crassa RNase N4
Mononucleotides
-
e.g. 2'-GMP, 3'-GMP, 5'-GMP, 3'-CMP, 2'(3')-UMP
additional information
-
fairly resistant to proteases, e.g. carboxypeptidase A, leucine aminopeptidase, trypsin, chymotrypsin
-
additional information
-
no effective inhibition of barnase by RNase inhibitor YrdF
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Nucleoside 5'-monophosphate
-
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.021
-
5'-OHGGGACAGUAUUUG-3'
-
R169K mutant; wild-type
0.00012
-
5'-pGGGACAGUAUUUG-3'
-
wild-type
0.0081
-
5'-pGGGACAGUAUUUG-3'
-
R169K mutant
0.052
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, N43H mutant
-
0.081
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, N43Q/N44G mutant
-
0.092
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, N43T mutant
-
0.17
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, Y45S mutant
-
0.19
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, N44H mutant
-
0.44
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, wild-type enzyme
-
0.61
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, N43I mutant
-
4
-
adenylyl-3',5'-cytidine
-
pH 6.0, 25C, N43G mutant
-
0.0268
-
GpA
-
-
0.069
-
GpC
-
RNase N1
0.238
-
GpC
-
RNase U1
0.435
-
GpC
-
RNase T1
0.9445
-
GpC
-
-
0.0291
-
GpG
-
-
0.0238
-
GpU
-
-
1670
-
guanosine-2',3'-cyclic monophosphate
-
pH 6.0, 25C, wild type enzyme
1730
-
guanosine-2',3'-cyclic monophosphate
-
pH 6.0, 25C, K41M mutant
2010
-
guanosine-2',3'-cyclic monophosphate
-
pH 6.0, 25C, K41T mutant
0.0581
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N43G mutant
0.0757
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, Q46_P47insQ mutant
0.0758
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N43I mutant
0.0772
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N43T mutant
0.135
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, wild-type enzyme
0.156
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N44H mutant
0.173
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N43H mutant
0.259
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N43Y mutant
0.361
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, Q46_Q47insP mutant
0.506
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, Q46_P47insP mutant
0.549
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, N43Q/N44G mutant
0.606
-
guanylyl-3',5'-cytidine
-
pH 6.0, 25C, Y45S mutant
5500
-
guanylyl-3',5'-uridine
-
pH 6.5, 25C, wild-type enzyme
5700
-
guanylyl-3',5'-uridine
-
pH 6.5, 25C, E74K mutant
0.151
-
poly(I)
-
25C, pH 6.2, wild-type enzyme
0.16
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79E; 25C, pH 6.5, wild-type enzyme
0.21
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79N
0.3
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79R
0.32
-
poly(I)
-
25C, pH 6.2, mutant enzyme E54Q
0.33
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79K
0.36
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79I
0.4
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79W
0.1
-
polyinosinic acid
-
pH 7.06, 25C, 5K mutant; pH 7.50, 25C, 5K mutant
0.11
-
polyinosinic acid
-
pH 8.10, 25C, 5K mutant
0.13
-
polyinosinic acid
-
pH 6.50, 25C, 5K mutant
0.14
-
polyinosinic acid
-
pH 6.50, 25C, 3K mutant; pH 7.06, 25C, 3K mutant; pH 8.50, 25C, 5K mutant
0.15
-
polyinosinic acid
-
pH 6.50, 25C, wild-type enzyme
0.16
-
polyinosinic acid
-
pH 6.05, 25C, wild-type enzyme
0.18
-
polyinosinic acid
-
pH 5.60, 25C, wild-type enzyme; pH 6.00, 25C, 3K mutant
0.19
-
polyinosinic acid
-
pH 7.06, 25C, wild-type enzyme; pH 7.48, 25C, 3K mutant
0.2
-
polyinosinic acid
-
pH 8.00, 25C, 3K mutant
0.21
-
polyinosinic acid
-
pH 5.05, 25C, wild-type enzyme
0.23
-
polyinosinic acid
-
pH 6.05, 25C, 5K mutant
0.24
-
polyinosinic acid
-
pH 4.47, 25C, wild-type enzyme; pH 8.48, 25C, 3K mutant
0.25
-
polyinosinic acid
-
pH 4.02, 25C, wild-type enzyme
0.26
-
polyinosinic acid
-
pH 5.50, 25C, 3K mutant; pH 7.50, 25C, wild-type enzyme
0.29
-
polyinosinic acid
-
pH 8.01, 25C, wild-type enzyme
0.33
-
polyinosinic acid
-
pH 5.00, 25C, 3K mutant
0.34
-
polyinosinic acid
-
pH 8.50, 25C, wild-type enzyme
0.47
-
polyinosinic acid
-
pH 5.60, 25C, 5K mutant
0.57
-
polyinosinic acid
-
pH 4.50, 25C, 3K mutant
0.64
-
polyinosinic acid
-
pH 5.04, 25C, 5K mutant
0.75
-
polyinosinic acid
-
pH 4.00, 25C, 3K mutant
1.1
-
polyinosinic acid
-
pH 4.47, 25C, 5K mutant
1510
-
polyinosinic acid
-
pH 6.5, 25C, wild-type enzyme
1530
-
polyinosinic acid
-
pH 6.0, 25C, wild type enzyme
1600
-
polyinosinic acid
-
pH 6.5, 25C, E41K mutant
1700
-
polyinosinic acid
-
pH 6.5, 25C, E74K mutant
2000
-
polyinosinic acid
-
pH 6.5, 25C, R65A mutant
2500
-
polyinosinic acid
-
pH 6.5, 25C, Q38A mutant
3040
-
polyinosinic acid
-
pH 6.0, 25C, K41M mutant
3060
-
polyinosinic acid
-
pH 6.0, 25C, K41T mutant
3100
-
polyinosinic acid
-
pH 6.5, 25C, E54Q mutant
0.321
-
RNA
-
pH 7.5, 25C, wild-type enzyme
0.433
-
RNA
-
pH 7.5, 25C, Y45S
0.698
-
RNA
-
pH 7.5, 25C, N43H
0.73
-
RNA
-
pH 7.5, 25C, N43G
0.961
-
RNA
-
pH 7.5, 25C, N43Y
1.55
-
RNA
-
pH 7.5, 25C, N43I
2.64
-
RNA
-
pH 7.5, 25C, N43T
4.56
-
RNA
-
pH 7.5, 25C, N43Q/N44G
6.94
-
RNA
-
pH 7.5, 25C, Q46PQ mutant
19.6
-
RNA
-
pH 7.5, 25C, N44H
1470
-
RNA
-
pH 7.5, 25C, Q46PP mutant
10000
-
guanylyl-3',5'-uridine
-
pH 6.5, 25C, E41K mutant
additional information
-
additional information
-
pH-dependence of Km
-
additional information
-
additional information
-
-
-
additional information
-
additional information
-
Kcat/Km values of 1000 mM-1 S-1, 0.458 mM-1 S-1, 7520 mM-1 S-1, and 8.65 mM-1 S-1 for wild type enzyme and Y38F, E58A, H92Q and F100A mutants, respectively, with GpU as substrate. Kcat/Km values of 11.3 mM-1 S-1, 15.1 mM-1 S-1, 0.153 mM-1 S-1, 27.4 mM-1 S-1, 0.117 mM-1 S-1, 1.38 mM-1 S-1 for wild type enzyme and Y38F, H40A, E58A, and F100A mutants, respectively, with SpGp(S)U as substrate. Kcat/Km values of 665 mM-1 S-1, 210 mM-1 S-1, 0.029 mM-1 S-1, 5.35 mM-1 S-1, 0.028 mM-1 S-1 and 0.508 mM-1 S-1 for wild type enzyme and Y38F, H40A, E58A, H92Q and F100A mutants, respectively, with RpGp(S)U as substrate
-
additional information
-
additional information
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, and 39 for mutant enzyne K41E/Y42F/N43R/Y45W/E46N/W59Y (RNase T1-R2) and mutant enzyme K41E/Y42F/N43R/Y45W/E46N (RNase RV)
-
additional information
-
additional information
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, 195 for mutant enzyme E46N and 137 for mutant enzyme E46N/Y45W
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
72
-
5'-OHGGGACAGUAUUUG-3'
-
wild-type
126
-
5'-pGGGACAGUAUUUG-3'
-
R169K mutant
180
-
5'-pGGGACAGUAUUUG-3'
-
wild-type
0.7
-
poly(I)
-
25C, pH 6.2, mutant enzyme E54Q
189
-
poly(I)
-
25C, pH 6.2, wild-type enzyme
220
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79E; 25C, pH 6.5, wild-type enzyme
280
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79N
410
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79I
430
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79K
460
-
poly(I)
-
25C, pH 6.5, mutant enzyme D79R; 25C, pH 6.5, mutant enzyme D79W
additional information
-
additional information
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, and 39 for mutant enzyne K41E/Y42F/N43R/Y45W/E46N/W59Y (RNase T1-R2) and mutant enzyme K41E/Y42F/N43R/Y45W/E46N (RNase RV)
-
additional information
-
additional information
-
kcat/Km(GpC)/kcat/Km(ApC) is 284000 for wild-type enzyme, 195 for mutant enzyme E46N and 137 for mutant enzyme E46N/Y45W
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.95
-
3'-N-hydroxyurea-3'-deoxythymidine-5'-phosphate
-
25C, pH 6.2, wild-type enzyme
1.15
-
3'-N-hydroxyurea-3'-deoxythymidine-5'-phosphate
-
25C, pH 6.2, mutant enzyme E54Q
0.000033
-
chitosan
-
poly(I) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
0.00008
-
chitosan
-
poly(A) * poly(U) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
0.00009
-
chitosan
-
poly(I) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
0.0001
-
chitosan
-
poly(I) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
0.00012
-
chitosan
-
poly(I) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
0.00021
-
chitosan
-
poly(I) * poly(C) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
0.00022
-
chitosan
-
poly(I) substrate, measurement is performed in 10 mM Tris, pH 7.0, containing 0.14 M NaCl and 0.1 mM EDTA
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.5
-
-
xanthosine 2',3'-cyclic phosphate
4.5
-
-
Aspergillus niger RNase II
5.5
-
-
assay at
6
-
-
Chalaropsis sp.
7
-
-
RNase N1
7.2
-
-
hydrolysis of guanosine 2',3'-cyclic phosphate
7.3
-
-
immobilized enzyme
7.4
-
-
assay at
7.5
-
-
RNA digestion
7.5
-
-
native enzyme
7.7
-
-
insolubilized enzyme
7.8
-
-
native enzyme
8
-
-
RNase U1
8
-
-
RNA cleavage assay
8.3
-
-
cleavage assay
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
6
-
3: 25% of activity maximum, 6: about 10% of activity maximum, Aspergillus niger RNase II
4
9
-
4: 10% (native), 20% (immobilized) of activity maximum, 9: 32% (native), 25% (immobilized) of activity maximum, 10: immobilized enzyme, some activity, native enzyme, inactive
4.5
8
-
4.5 and 8: Chalaropsis sp., similar enzyme
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
RNA cleavage assay
37
-
-
cleavage assay
43
-
-
immobilized enzyme
45
-
-
native enzyme
50
-
-
Aspergillus niger RNase II
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
55
-
20C: 18% of activity maximum, 55C: 15% of activity maximum, inactive at 60C, native enzyme
38
80
-
38C: 55% of activity maximum, 80C: 40% of activity maximum, immobilized enzyme
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
-
-
isoelectric focusing, wild-type enzyme
4.6
4.7
-
calculated from titration curve, 5K mutant
4.9
-
-
calculated from titration curve, 2K mutant; isoelectric focusing, 2K mutant
5.5
-
-
isoelectric focusing, 5K mutant
6
-
-
isoelectric focusing, 3K mutant
6.9
-
-
calculated from titration curve, 4K mutant; isoelectric focusing, 4K mutant
7.1
-
-
calculated from titration curve, 3K mutant
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
Taka-diastase, commercial product of Aspergillus oryzae
Manually annotated by BRENDA team
additional information
-
binase is not produced by Bacillus pumilus when bacteria are grown on medium containing an inorganic source of nitrogen. Expression of binase is possible only in Spo0A-OFF cells
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
in contrast to the wild-type strain in which the inactivated RNase T1-GFP fusion protein is localized at the vacuole only under cold stress or nitrogen starvation, the inactivated RNase T1-GFP fusion protein expressed in the rns4 mutant is localized at the ER and vacuole, both under normal growth conditions and upon ambient stress conditions
Manually annotated by BRENDA team
additional information
-
about 3040% of barnase can be stored inside the cells for up to 90 min in complex with inhibitor barstar
-
Manually annotated by BRENDA team
additional information
-
99% of binase is present outside of the cell and only below 1% is localized intracellularly
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Erwinia carotovora subsp. atroseptica
Erwinia carotovora subsp. atroseptica
Erwinia carotovora subsp. atroseptica
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
10000
-
-
Neurospora crassa, RNase N4, gel filtration, SDS-PAGE
11000
-
-
Ustilago sphaerogena, amino acid composition
11000
-
-
Fusarium moniliforme, gel filtration
11090
-
-
Aspergillus oryzae, amino acid analysis
11840
-
-
Chalaropsis sp., amino acid composition
12000
-
-
Chalaropsis sp., low speed equilibrium ultracentrifugation
13000
-
-
Aspergillus niger, RNase II, gel filtration
53000
-
-
fusion protein of the antiferritin antibody VL domain and barnase, gel filtration
71100
-
-
apparent molecular mass of the monomer, determined by sedimentation equilibrium analysis
79000
-
-
determined by SDS-PAGE, N-terminally His-tagged form
325000
-
-
tetramer, determined by gel-filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
-
2 * 26000, fusion protein of the antiferritin antibody VL domain and barnase, SDS-PAGE
monomer
-
primary structure, Aspergillus oryzae, 1 * 11085
monomer
-
Chalaropsis sp.
monomer
-
1 * 12382, sequence calculation
monomer
-
1 * 12213, sequence calculation
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no glycoprotein
-
Chalaropsis sp.
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Aspergillus oryzae
-
crystal structure of RNase T1-T2 (K41E/Y42F/N43R/Y45W/E46N/W59Y), hanging drop vapor diffusion method, 2.1 A resolution
-
hanging drop vapor diffusion method, crystallization of RNase T1 variant RV (mutant RNase T1 K41E/Y42F/N43R/Y45W/E46N)
-
macroseeding
-
two crystal forms obtained, the classic crystal form I, and the crystals of form II, obtained by repeated seeding
-
the X-ray coordinates of barnase for this study are obtained from the Protein Data Bank, PDB code 1rnb
-
Streptomyces aureofaciens
-
Neurospora crassa RNase N1
-
the crystal structures of RNase NT in complex with either 5'-AMP, 5'-GMP, or 2'-UMP are determined at 1.8 A resolutions by molecular replacement
-
hanging-drop vapor diffusion
-
mutant enzyme Q94K, hanging-drop vapor-diffusion method at 18C
-
RNase Sa-3'IMP complex
-
structures of two crystal forms (I and II) of ribonuclease Sa2 are presented at 1.8 A and 1.5 A resolution. Vapour diffusion from a solution of 1.0% protein by weight in 0.1 M phosphate buffer at pH 7.2 and room temperature with 40% saturated ammonium sulfate as precipitant
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.2
-
-
37C, 22 h
3
9
-
stable
4
10
-
37C, 33 h, immobilized enzyme, highest stability at pH 7, at pH 1.0 and 10.0: about 60% of activity maximum
4.5
-
-
maximal conformational stability
7
-
-
-20C, several months, 4C, several weeks, 37C, 22 h
7.5
-
-
25C, 40 h
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
50
-
-
30 min, Neurospora crassa RNase N4
60
-
-
30 min, 60% loss of activity, Neurospora crassa RNase N4
70
-
-
30 min, 90% loss of activity, Neurospora crassa RNase N4
100
-
-
10 min, pH 6; above pH 9 unstable
100
-
-
30 min, immobilized enzyme: 5% loss of activity, native enzyme: about 60% loss of activity
additional information
-
-
the reversibility of the thermal denaturation is generally between 90% and 95%. Q38A and E74K mutants are slightly more stable than the wild-type enzyme. E41K, E54A and R65A mutants are less stable than the wild-type enzyme
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Conformational stability
-
Guanidine hydrochloride, pH-dependence of denaturation
-
Immobilized enzyme, heat- and pH-stability
-
Insolubilized enzyme
-
MgCl2 increases conformational stability
-
Na2HPO4 increases conformational stability
-
NaCl, protects from thermal denaturation, increases transition temperature, suppresses unfolding of enzyme by a denaturant, urea, increases conformational stability
-
Proteases, fairly resistant to proteases: carboxypeptidase A, leucine aminopeptidase, trypsin, chymotrypsin <1>, resistant to trypsin and chymotrypsin, sensitive to pepsin
-
Spermine stabilizes
-
the enzyme is remarkably stable at high salt concentrations
-
Urea, 8 M, stable
-
Urea, pH-dependence of urea denaturation
-
NaCl and low concentrations of guanidinium chloride stabilize RNase Sa, conformational stability is studied by NMR-monitored hydrogen exchange
P05798
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Photooxidation, in presence of Methylene Blue, riboflavine or Rose Bengal inactivation
-
24191
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, pH 7, several months
-
4C, pH 6, insolubilized enzyme, 5 weeks
-
4C, pH 7, several weeks
-
-80C, HEPES buffer, pH 7.5, 0.1 mM EDTA, 1 mM DTT, 50 mM NaCl
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
similar enzyme, partial
-
anion-exchange chromatography and gel filtration
-
diethylaminoethylcellulose chromatography
-
gel filtration
-
ion-exchange chromatography and gel filtration
-
on porous glass affinity adsorbent
-
RNase T1 variants E46N and Y45W/E46N
-
using a Super Q Toyopearl column and ion-exchange chromatography on a column of resource Q
-
fusion protein of the antiferritin antibody VL domain and barnase
-
similar enzyme, partial
-
using Ni-NTA spin columns
-
usung a HisTrap HP column
-
similar enzyme
-
ion-exchange chromatography and gel filtration
-
using Ni2+-nitrilotriacetic acid column chromatography
-
similar enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Epicurian coli
-
expression in Escherichia coli
-
expression in Saccharomyces cerevisiae
-
overexpression in Saccharomyces cerevisiae. Characterization of an rns4/vps32 mutation in the RNase T1 expression-sensitive strain of Saccharomyces cerevisiae. The rns4 mutant is sensitive to both RNase T1 and ambient stress. In contrast to the wild-type strain in which the inactivated RNase T1-GFP fusion protein is localized at the vacuole only under cold stress or nitrogen starvation, the inactivated RNase T1-GFP fusion protein expressed in the rns4 mutant is localized at the ER and vacuole, both under normal growth conditions and upon ambient stress conditions
-
fusion of the antiferritin antibody VL domain to barnase and expression in Escherichia coli
-
genetic organization, low level expression of barnase by recombinant Bacillus subtilis strains bearing phoPR-null mutations
-
the secondary structure unit mutant S2354 is expressed in Escherichia coli only when His102 is substituted by alanine (H102A)
-
expression in Escherichia coli BL21 carrying plasmid pGEMGX1/ent/Bi as a homogeneous protein
-
genetic organization, production of binase by recombinant Bacillus subtilis strains bearing phoPR-null mutations is impossible
-
chemically synthesized gene, mutant Gln58Asp, expressed in Escherichia coli host
-
for expression in Escherichia coli BL21DE3-Gold cells
-
gene rng, overexpression of wild-type rng and of mutant rngs in rne-1 or rneD1018 alleles mutant strains
-
a 1.8 kb DNA fragment, encoding the C-terminal portion of the MycRne polypeptide, residues 332-953, is cloned into p6HisF-11d for expression in Escherichia coli BL21-CodonPlusDE3-RIL cells
-
overexpression in Neurospora crassa, different promoters are tested, the most promising promoter for recombinant expression is the cfp promoter
P09646
into the pGEM-T vector for transformation of Escherichia coli JM109 cells, into pPIC9K for expression in Pichia pastoris
-
into the pQE30 vector for expression in Escherichia coli M15 cells
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
upregulation of the KCa channels and the apoptotic p53 genes, downregulation of the antiapoptotic bcl-2 gene under binase action
-
low concentrations of extracellular inorganic phosphate induce the expression of the binase gene
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E41K
-
with almost the same activity as wild-type enzyme. Glu41 is not involved in substrate binding
E41K/D17K
-
2K mutant
E41K/D17K/D1K
-
3K mutant
E41K/D17K/D1K/D25K
-
4K mutant
E41K/D17K/D1K/D25K/E74K
-
5K mutant
E46N
-
mutant shows an improved ApC/GpC preference with a 1450fold increase in comparison to wild-type activity
E54Q
-
dramatically less active than the wild-type enzyme
E58A
-
less active than the wild type enzyme
E58D
-
with dramatically decreased activity
E74K
-
less active than the wild-type enzyme due to a change in the orientation of the catalytic groups
F100A
-
less active than the wild type enzyme
H40A
-
less active than the wild type enzyme
H85Q
-
dramatically less active than the wild-type enzyme
H92Q
-
less active than the wild type enzyme
K41E/Y42F/N43R/Y45W/E46N/W59Y
-
RNase T1-R2 contains the additional mutation W59Y compared to RNase T1-RV variant. RNase T1-RV is a variant where the specificity is changed from guanine to purine, accompanied with a reduced activity. The additional mutation W59Y increases the activity in comparison to variant RV to 425%
K41M
-
mutant with identical catalytic properties as the wild type enzyme with guanosine-2',3'-cyclic monophosphate as substrate, but less active with polyinosinic acid as substrate
N43G
-
as active as the wild type enzyme
N43H
-
as active as the wild type enzyme
N43I
-
as active as the wild type enzyme
N43Q/N44G
-
less active than the wild type enzyme
N43T
-
as active as the wild type enzyme
N43Y
-
as active as the wild type enzyme
N44A
-
with dramatically decreased activity
N44H
-
less active than the wild type enzyme
Q38A
-
less active than the wild-type enzyme
Q46_G47insL
-
mutant with a Leu insertion in 47, less active than the wild type enzyme
Q46_N47insQ
-
mutant with a Glu insertion in 47, less active than the wild type enzyme
Q46_P47insP
-
mutant with a Pro insertion in 47, less active than the wild type enzyme
Q46_P47insQ
-
mutant with a Glu insertion in 47, as active as the wild type enzyme
Q46_P47insY
-
mutant with a Tyr insertion in 47, less active than the wild type enzyme
Q46_Q47insP
-
mutant with a Pro insertion in 47, less active than the wild type enzyme
Q46_R47insR
-
mutant with an Arg insertion in 47, less active than the wild type enzyme
Q46_S47insG
-
mutant with a Gly insertion in 47, less active than the wild type enzyme
Q46_S47insS
-
mutant with a Ser insertion in 47, less active than the wild type enzyme
Q46_T47insS
-
mutant with a Ser insertion in 47, less active than the wild type enzyme
R65A
-
dramatically less active than the wild-type enzyme
R77K
-
with dramatically decreased activity
V16A
-
considerably less stable than the wild-type enzyme
V16C
-
considerably less stable than the wild-type enzyme
V16S
-
considerably less stable than the wild-type enzyme
V16T
-
considerably less stable than the wild-type enzyme
V78A
-
considerably less stable than the wild-type enzyme
V78C
-
considerably less stable than the wild-type enzyme
V78S
-
considerably less stable than the wild-type enzyme
V78T
-
considerably less stable than the wild-type enzyme
V89A
-
considerably less stable than the wild-type enzyme
V89C
-
considerably less stable than the wild-type enzyme
V89S
-
considerably less stable than the wild-type enzyme
V89T
-
considerably less stable than the wild-type enzyme
Y38F
-
less active than the wild type enzyme
Y45S
-
as active as the wild type enzyme
Y45W/E46N
-
mutant shows an improved ApC/GpC preference with a 2100fold increase in comparison to wild-type activity
D54A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
D54A/E60A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
E60A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
H102A
-
the secondary structure unit mutant S2354 is expressed in Escherichia coli only when His102 is substituted by alanine (H102A). The mutant S2354 102A has secondary and tertiary structures and unfolds in a cooperative manner during urea-induced unfolding experiments. S2354H102A interacts with other barnase mutants to hydrolyze RNA
K27A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
K66A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
R59A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
R83Q
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
R87A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
D303N
-
mutant with amino acid substitution in the binding pocket, inactive enzyme
R169A
-
mutant with amino acid substitution in the binding pocket, inactive enzyme
R169K
-
mutant with amino acid substitution in the binding pocket
T170V
-
mutant with amino acid substitution in the binding pocket
V128A
-
mutant with amino acid substitution in the binding pocket
C196T/C603T
-
double mutant to avoid restriction enzymatic scission
D1W
-
the mutant is studied in concentrated urea and GdnHCl solution with their disulfide bond broken, the results show that long-range effects in a denaturated protein can significantly effect the fluorescence properties
D33A
-
Tm-value at pH 7.0 in Mops buffer is 16 C lower than wild-type value. The stability of the mutant enzyme is 6 kcal/mol less than wild-type RNase Sa
D79A
-
Tm-value at pH 7.0 in Mops buffer is 9.2 C higher than wild-type value. The stability of the mutant enzyme is 3.3 kcal/mol less than wild-type RNase Sa
D79E
-
Tm-value at pH 7.0 in Mops buffer is 0.8 C lower than wild-type value. kcat/Km is identical to wild-type value
D79F
-
Tm-value at pH 7.0 in Mops buffer is 9.9 C higher than wild-type value
D79H
-
Tm-value at pH 7.0 in Mops buffer is 5.6 C higher than wild-type value
D79I
-
Tm-value at pH 7.0 in Mops buffer is 9.6 C higher than wild-type value. kcat/Km is 1.3fold lower than wild-type value
D79K
-
Tm-value at pH 7.0 in Mops buffer is 7.6 C higher than wild-type value. kcat/Km is 1.1fold lower than wild-type value
D79L
-
Tm-value at pH 7.0 in Mops buffer is 8.7 C higher than wild-type value
D79N
-
Tm-value at pH 7.0 in Mops buffer is 5.5 C higher than wild-type value. kcat/Km is 1.1fold lower than wild-type value
D79R
-
Tm-value at pH 7.0 in Mops buffer is 9.0 C higher than wild-type value. kcat/Km is 1.1fold higher than wild-type value
D79W
-
Tm-value at pH 7.0 in Mops buffer is 7.6 C higher than wild-type value. kcat/Km is 1.2fold lower than wild-type value
D79Y
-
Tm-value at pH 7.0 in Mops buffer is 9.6 C higher than wild-type value
Q94K
-
Tm-value at pH 7.0 in Mops buffer is 0.8 C higher than wild-type value. Crystal structure shows that the amino group of the Lys forms a hydrogen-bonded ion pair with the carboxyl group of Asp79. The stability of the mutant is about the same as the wild-type at pH 3, where Asp79 is uncharged, but 1 kcal/mol greater than that of wild-type RNase Sa at pH 8.5, where Asp79 is charged
T76W
-
the mutant is studied in concentrated urea and GdnHCl solution with their disulfide bond broken, the results show that long-range effects in a denaturated protein can significantly effect the fluorescence properties
Y52W
-
the mutant is studied in concentrated urea and GdnHCl solution with their disulfide bond broken, the results show that long-range effects in a denaturated protein can significantly effect the fluorescence properties
Y55W
-
the mutant is studied in concentrated urea and GdnHCl solution with their disulfide bond broken, the results show that long-range effects in a denaturated protein can significantly effect the fluorescence properties
Y81W
-
the mutant is studied in concentrated urea and GdnHCl solution with their disulfide bond broken, the results show that long-range effects in a denaturated protein can significantly effect the fluorescence properties
K41T
-
mutant with identical catalytic properties as the wild type enzyme with guanosine-2',3'-cyclic monophosphate as substrate, but less active with polyinosinic acid as substrate
additional information
-
two extended replica-exchange molecular dynamics simulations on RNase T1, each with different histidine protonation states, corresponding to pH 6 and pH8. At high pH, the appearance of partially unfolded states is evident. This pH-induced destabilization originates from increased global repulsion as well as reduced local favorable electrostatic interactions and reduced H-bonding strength of His27, His40, and His92. At high pH, alternative tryptophan rotamers appear and are linked to a distorted environment of the tryptophan, which also acts as a separate source of ground-state heterogeneity
K66A/D54A
-
mutant is modeled in silico for gaining insight into modulations of diffusional association behavior, which is reflected in the association rate
additional information
-
fusion of the antiferritin antibody VL domain to barnase results in enhanced solubility and altered pH stability
D346N
-
mutant with amino acid substitution in the binding pocket
additional information
-
two distinct spontaneously derived single amino acid substitutions within the predicted RNase H domain of RNase G, generate the rng-219 and rng-248 alleles, which complement the rne mutant strains. Domain swaps between RNase E and RNase G generate proteins that do not complement RNase E deficiency
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
salts induce chain folding of disulfide-reduced and modified RNase T1 into native conformation, enzyme activity is not restored
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
drug development
-
bacterial RNases are promising tools for the development of anticancer drugs. Binase affects the total amount of intracellular RNA and the expression of proapoptotic and antiapoptotic mRNAs
medicine
-
RNase inhibitors appear to be promising for therapy of cancer
biotechnology
-
the RNase G mutation could be applied in the breeding of producer strains of pyruvate and its derivatives such as valine
medicine
-
RNase inhibitors appear to be promising for therapy of cancer
medicine
-
RNase inhibitors appear to be promising for therapy of cancer
additional information
-
acquisition of male, female or bisexual sterility by transgenic plants with the help of barnase. The enzyme is useful in medical research, overview
medicine
-
RNase inhibitors appear to be promising for therapy of cancer
additional information
-
the enzyme is useful in medical research, overview