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Information on EC 3.1.21.B3 - mismatch-specific endonuclease and Organism(s) Thermococcus kodakarensis and UniProt Accession Q5JER9

for references in articles please use BRENDA:EC3.1.21.B3
preliminary BRENDA-supplied EC number
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Thermococcus kodakarensis
UNIPROT: Q5JER9
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The taxonomic range for the selected organisms is: Thermococcus kodakarensis
The expected taxonomic range for this enzyme is: Thermococcus kodakarensis
Reaction Schemes
endonucleolytic cleavage of both strands of double-stranded DNA into 5'-protruding forms, with the mismatched base pair in the central position. The enzyme cleaves G/T, G/G, T/T, T/C and A/G mismatches, with a preference for G/T, G/G and T/T. It cleaves the third phosphodiester bond on the 5' side of the mismatched base in both strands, to produce a cohesive end with a five-nucleotide long 5'-protrusion, leaving 5'-phosphate and 3'-hydroxyl termini.
Synonyms
mismatch-specific endonuclease, endonuclease nucs, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
DNA + H2O
?
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
DNA + H2O
?
show the reaction diagram
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
the optimal temperature of the reaction is around 60°C
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
28595
monomer, calculated from sequence
60200
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
2 * 28595, calculated from sequence, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
the structures of TkoEndoMS in complex with dsDNA substrates and in unbound (apo) form are determined at a maximum 2.4 A resolution with X-ray crystallography. The crystals of apo TkoEndoMS are grown in a stock solution (50 mM Tris-HCl [pH 8.0], 0.1 mM EDTA, 0.5 mM DDT, 10% [w/v] glycerol, 0.6 M NaCl, and 445 mM TkoEndoMS) stored at 4°C. The DNA cleavage reaction of the enzyme in the crystals is inhibited by replacing Asp165, one of the putative catalytic sites, with Ala
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D165A
the DNA cleavage reaction of the enzyme in the crystals is inhibited by replacing Asp165, one of the putative catalytic sites, with Ala
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
80
80% of the activity remains after 30 min at 80°C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
activity is distinctly salt-resistant in vitro
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli BL21
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
molecular biology
the thermostable mismatch-specific endonuclease will be applicable to genetic engineering techniques
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ishino, S.; Nishi, Y.; Oda, S.; Uemori, T.; Sagara, T.; Takatsu, N.; Yamagami, T.; Shirai, T.; Ishino, Y.
Identification of a mismatch-specific endonuclease in hyperthermophilic Archaea
Nucleic Acids Res.
44
2977-2986
2016
Thermococcus kodakarensis (Q5JER9), Thermococcus kodakarensis, Thermococcus kodakarensis ATCC BAA-918 (Q5JER9)
Manually annotated by BRENDA team
Nakae, S.; Hijikata, A.; Tsuji, T.; Yonezawa, K.; Kouyama, K.I.; Mayanagi, K.; Ishino, S.; Ishino, Y.; Shirai, T.
Structure of the EndoMS-DNA complex as mismatch restriction endonuclease
Structure
24
1960-1971
2016
Thermococcus kodakarensis (Q5JER9), Thermococcus kodakarensis ATCC BAA-918 (Q5JER9)
Manually annotated by BRENDA team