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3.1.31.1: micrococcal nuclease

This is an abbreviated version!
For detailed information about micrococcal nuclease, go to the full flat file.

Word Map on EC 3.1.31.1

Reaction

endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotide end-products =

Synonyms

AtTSN1, AtTSN2, cTSN, EC 3.1.4.7, Epstein-Barr virusencoded transcription factor 2 co-activator p100, HsTSN, micrococcal DNase, micrococcal endonuclease, Micrococcal nuclease, MN, MN ase, MNase, Nuc, NUC1, Nuc2, nucB, nuclease 8V, nuclease T, nuclease T', nuclease, micrococcal, nuclease, staphylococcal, NucM, P100, PaTSN, PfTSN, ribonucleate (deoxyribo-nucleate) 3'-nucleotidohydrolase, ribonucleate (deoxyribonucleate) 3'-nucleotidohydrolase, S. aureus nuclease, SNA, snake venom phosphodiesterase, SNase, SNAseR, SND1, spleen endonuclease, spleen phosphodiesterase, staph nuclease, staphylococcal nuclease, Staphylococcal nuclease A, Staphylococcal nuclease domain containing-1, staphylococcal nuclease domain-containing 1, staphylococcal nuclease domain-containing protein 1, staphylococcus aureus nuclease, staphylococcus aureus nuclease B, Staphylococcus aureus nuclease homologue, thermonuclease, TNase, TSN, tudor staphylococcal nuclease, Tudor-SN, Tudor-staphylococcal nuclease

ECTree

     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.31 Endoribonucleases that are active with either ribo- or deoxyribonucleic acids and produce 3'-phosphomonoesters
                3.1.31.1 micrococcal nuclease

Engineering

Engineering on EC 3.1.31.1 - micrococcal nuclease

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D790E
by site-directed mutagenesis, proteolysis of Tudor staphylococcal nuclease does not occur either when the P1 position of the DAVD motif is mutated or after treatment with the pan-caspase inhibitor zVAD-fluoromethylketone, expression of mutant under normal conditions enhances cell proliferation in both cancer (HeLa) and non-cancer (HEK-293) cells compared with mock- and wild-type Tudor staphylococcal nuclease-transfected samples, under camptothecin-induced apoptosis, expression of Tudor staphylococcal nuclease mutant results in a 35% increment in viable HeLa cells, suggesting that caspase-mediated proteolysis of enzyme is important for the progression of apoptosis
DELTA140-149
-
deletion of the 10 C-terminal residues, mutant proteins are in a non-native or disordered state under physiological conditions, folding is induced by addition of an inhibitor or substrate
F34A
-
site-directed mutagenesis
G79S/H124LC80-C116
-
effects on the stability and conformation of the folded protein
H124LC77-C118
-
effects on the stability and conformation of the folded protein
H124LC79-C118
-
effects on the stability and conformation of the folded protein
H124LC80-C116
-
effects on the stability and conformation of the folded protein
I92A
-
site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
INS33A34
-
insertion of an alanine between residues 33 and 34, mutant proteins are in a non-native or disordered state under physiological conditions, folding is induced by addition of an inhibitor or substrate
K45C
-
insertion of a cysteine to enable labeling with thiol reactive ligands, e.g. 5,5'-dithiobis-2-nitrobenzoic acid, CD-spectra of wild type enzyme, mutant and mutant with 5,5'-dithiobis-2-nitrobenzoic acid label indicate, that the protein have very similar secondary structures
L103A
-
site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
L125A
-
site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
L25A
-
site-directed mutagenesis
L36A
-
site-directed mutagenesis
L38A
-
site-directed mutagenesis
P117G,/H124L/S128A
-
site-directed mutagenesis, a highly stable triple mutant
P117G/H124L/S128A
-
site-directed mutagenesis
P11A/P31A/P42A/P47T/P56A/P117G
proline free mutant, conformationally different from wild type protein, 1.4% of wild type activity
P47G/P117G/H124L/W140H
-
tryptophan-free mutant used for the insertion of a unique tryptophan at positions 15, 27, 61, 76, 91, 102, and 121, mutant enzymes used to study the enzyme folding kinetics, variants are destabilized but maintain the ability to refold in the native-like structure
T62C
-
designed for the insertion of a cysteine reactive label
T62P
highly destabilized variant of enzyme, exists in the unfolded state over a wide pH-range, can be fully refolded to the native folding by addition of osmolytes
V23A
-
site-directed mutagenesis
V66A
-
site-directed mutagenesis, the mutant shows similar global stability like the wild-type enzyme
V66F/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66G/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66N/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66Q/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66S/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66T/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V66Y/P117G,/H124L/S128A
-
site-directed mutagenesis of the highly stable triple mutant P117G,/H124L/S128A, thermodynamic stability during guanidine hydrochloride denaturation of mutants is compared
V74A
-
site-directed mutagenesis
A128S
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
A1C/Q149C
-
SNase double mutant, N- and C-terminal residues replaced by cysteine, constructed from the plasmid (pMT7-SN) of wild-type SNase using the Kunkel method, can form disulfide bond
A90S
-
pH-value, at which the acid-denaturation is half completed is 4.19, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.0 for the mutant enzyme compared to 1.8 for wild-type enzyme
D143G
-
Tm value for mutant enzyme is 50.53°C, compared to 50.98°C for wild-type enzyme
D143K
-
charge reversal
D143N
-
charge reversal
D146G
-
Tm value for mutant enzyme is 50.99°C, compared to 50.98°C for wild-type enzyme
D19G
-
Tm value for mutant enzyme is 52.06°C, compared to 50.98°C for wild-type enzyme
D19K
-
charge reversal
D19L
-
charge reversal
D21G
-
Tm value for mutant enzyme is 53.74°C, compared to 50.98°C for wild-type enzyme
D21K
-
charge reversal
D21N
-
charge reversal
D21N/T33V/T41I/S59A/P117G/A128A
-
hyperstable engineered form of staphylococcal nuclease (SNase)
D40G
-
Tm value for mutant enzyme is 50.44°C, compared to 50.98°C for wild-type enzyme
D77A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
D77K
-
charge reversal
D83G
-
Tm value for mutant enzyme is 37.21°C, compared to 50.98°C for wild-type enzyme
D95G
-
Tm value for mutant enzyme is 37.38°C, compared to 50.98°C for wild-type enzyme
DELTA1-139
-
mutant lacks tertiary structure, fluorescence of the mutant is much lower than that of the wild-type enzyme
DELTA1-141
-
intact tertiary conformation, melting point is nearly identical to wild-type enzyme
E101A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine shortrange effects on His124, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
E101G
-
Tm value for mutant enzyme is 43.04°C, compared to 50.98°C for wild-type enzyme
E10G
-
Tm value for mutant enzyme is 43.8°C, compared to 50.98°C for wild-type enzyme
E10K
-
charge reversal
E10Q
-
charge reversal
E122G
-
Tm value for mutant enzyme is 44.12°C, compared to 50.98°C for wild-type enzyme
E122K
-
charge reversal
E122Q
-
charge reversal
E129G
-
Tm value for mutant enzyme is 34.59°C, compared to 50.98°C for wild-type enzyme
E135G
-
Tm value for mutant enzyme is 44.54°C, compared to 50.98°C for wild-type enzyme
E135K
-
charge reversal
E135Q
-
charge neutralization
E142G
-
Tm value for mutant enzyme is 49.41°C, compared to 50.98°C for wild-type enzyme
E43G
-
Tm value for mutant enzyme is 54.99°C, compared to 50.98°C for wild-type enzyme
E52G
-
Tm value for mutant enzyme is 52.1°C, compared to 50.98°C for wild-type enzyme
E57G
-
Tm value for mutant enzyme is 46.6°C, compared to 50.98°C for wild-type enzyme
E57K
-
charge reversal
E57Q
-
charge reversal
E67G
-
Tm value for mutant enzyme is 46.53°C, compared to 50.98°C for wild-type enzyme
E67Q
-
charge reversal
E73A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
E73G/D77G
-
loss of thermal stabilty of 47% relative to the wild-type protein
E73G/E75G
-
loss of thermal stabilty of 59% relative to the wild-type protein
E73G/E75G/D77G
-
loss of thermal stabilty of 65% relative to the wild-type protein
E73K
-
charge reversal
E73Q
-
charge reversal
E75A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy), determination of tautomeric states of His121 and His124, pH near pI
E75G
-
Tm value for mutant enzyme is 36.99°C, compared to 50.98°C for wild-type enzyme
E75G/D77G
-
loss of thermal stabilty of 58% relative to the wild-type protein
E75K
-
charge reversal
F34W/W140F
-
characterized by far and near UV CD, gel filtration, ANS-binding fluorescence, enzymatic parameters are similar to those of the wild type, similar substrate affinity to the wild type enzyme
F61W/W140A
-
the mutant shows reduced activity with higher Michaelis-Menten constants, Km, and lower maximum reaction rate compared to the wild-type enzyme, the mutant also shows a more rapid loss of secondary and tertiary structure by Gdn-HCl unfolding than the wild-type enzyme
F76W/W140H
-
mutation causes decrease in thermal stability
G20A
-
2% of wild-type activity. Km(Ca) is almost 20fold higher than the wild-type enzyme. Denaturation midpoint for the mutant enzyme is 1.5 M urea compared to 2.0 M urea for the wild-type enzyme
G20I
-
0.21% of wild-type activity. Km(Ca) is almost 50fold higher than the wild-type enzyme. Denaturation midpoint for mutant enzyme is 0.82 M urea compared to 2.0 M urea for the wild-type enzyme
G20V
-
0.45% of wild-type activity. Km(Ca) is almost 20fold higher than the wild-type enzyme. Denaturation midpoint for the mutynt enzyme is 1.1 M urea compared to 2.0 M urea for the wild-type enzyme
G50F/V51N/P117G/H124L/S128A/DELTA44-49
-
hyperstable, acid-resistant mutant form of enzyme known as delta+PHS
G79S
-
pH-value, at which the acid-denaturation is half completed is 4.39, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.4 for the mutant enzyme compared to 1.8 for wild-type enzyme
G88V
-
pH-value, at which the acid-denaturation is half completed is 3.57, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 3.0 for the mutant enzyme compared to 1.8 for wild-type enzyme
H124E
-
charge reversal
H124L
-
pH-value, at which the acid-denaturation is half completed is 2.98, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 2.8 for the mutant enzyme compared to 1.8 for wild-type enzyme
H124Q
-
charge neutralization
I92E
-
wild-type enzyme exhibits a broad range of pH-independence from pH 4.5 to pH 9, mutant enzyme exhibits pronounced pH-dependence with a maximal stability at pH 4.9
I92K
-
wild-type enzyme exhibits a broad range of pH-independence from pH 4.5 to pH 9.0, mutant enzyme exhibits pronounced pH-dependence with a maximal stability at pH 9.8
K127A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine shortrange effects on His124, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
K127E
-
charge reversal
K127Q
-
charge neutralization
K133A
-
intact tertiary conformation, melting point is 4.6°C lower than that of the wild-type enzyme
K133Q
-
charge neutralization
K28C/H124C
-
generated by site-directed mutagenesis, native and non-native conformations are observed, and the non-native conformation expands with increasing guanidinium hydrochloride concentrations, the non-native chains of the derivative exhibits different changes of persistence length at higher guanidinium hydrochloride concentrations, suggesting a subdomain-specific collapse of the denatured state of SNase, this local chain specific collapse is likely to play a role in modulating the formation of early intermediate during protein folding
K28C/K97C
-
generated by site-directed mutagenesis, native and non-native conformations are observed, and the non-native conformation expands with increasing guanidinium hydrochloride concentrations, the non-native chains of the derivative exhibits different changes of persistence length at higher guanidinium hydrochloride concentrations, suggesting a subdomain-specific collapse of the denatured state of SNase, this local chain specific collapse is likely to play a role in modulating the formation of early intermediate during protein folding
K28E
-
charge reversal
K28Q
-
charge neutralization
K48E
-
charge reversal
K48Q
-
charge neutralization
K63E
-
charge reversal
K63Q
-
charge neutralization
K64E
-
charge reversal
K64Q
-
charge neutralization
K70E
-
charge reversal
K70Q
-
charge neutralization
K78E
-
charge reversal
K78Q
-
charge neutralization
K84E
-
charge reversal
K84Q
-
charge neutralization
K97E
-
charge reversal
K97Q
-
charge neutralization
K9A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to study inductive effects and longer-range interactions between elements of the network, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
L25A
-
pH-value, at which the acid-denaturation is half completed is 4.15, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.2 for the mutant enzyme compared to 1.8 for wild-type enzyme
S28C
-
this mutant contains a five-amino acid type I beta-turn from concanavalin A in place of residues 27-30 of SNase
V66D/P117G/H124L/S128A
-
production by site-directed mutagenesis, pka value shifts to 7.79 and, after chemical denaturation, to 8.05
V66D/P117G/H124L/S128A/G50F/V51N/DELTA44-49
-
production by site-directed mutagenesis, pka value shifts to 8.95 and, after chemical denaturation, to 8.73
V66E/P117G/H124L/S128A
-
production by site-directed mutagenesis, pka value shifts to 8.80 and, after chemical denaturation, to 8.99
V66E/P117G/H124L/S128A/G50F/V51N/DELTA44-49
-
production by site-directed mutagenesis, pka value shifts to 9.07 and, after chemical denaturation, to 8.80
V66K/P117G/H124L/S128A
-
production by site-directed mutagenesis, pka value shifts to 6.35
V66K/P117G/H124L/S128A/G50F/V51N/DELTA44-49
-
production by site-directed mutagenesis, pka value shifts to 5.63 and, after chemical denaturation, to 5.83
V66L
-
pH-value, at which the acid-denaturation is half completed is 3.36, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 2.6 for the mutant enzyme compared to 1.8 for wild-type enzyme
V66L/G79S/G88V
-
pH-value, at which the acid-denaturation is half completed is 3.67, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.1 for the mutant enzyme compared to 1.8 for wild-type enzyme
V66L/G88V
-
pH-value, at which the acid-denaturation is half completed is 3.42, compared to pH 3.76 for wild-type enzyme. The apparent number of protons which trigger the denaturation and are taken up by the protein upon denaturation is 1.6 for the mutant enzyme compared to 1.8 for wild-type enzyme
W140A
W140C
-
DNA hydrolysis activity is 75% of wild-type activity
W140D
-
DNA hydrolysis activity is 65% of wild-type activity
W140E
-
DNA hydrolysis activity is 65% of wild-type activity
W140F
W140G
-
DNA hydrolysis activity is 75% of wild-type activity
W140H
W140I
-
DNA hydrolysis activity is 70% of wild-type activity
W140K
-
DNA hydrolysis activity is 70% of wild-type activity
W140L
W140M
-
DNA hydrolysis activity is 70% of wild-type activity
W140N
-
DNA hydrolysis activity is 75% of wild-type activity
W140P
-
DNA hydrolysis activity is 55% of wild-type activity
W140Q
-
DNA hydrolysis activity is 75% of wild-type activity
W140R
-
DNA hydrolysis activity is 75% of wild-type activity
W140S
-
DNA hydrolysis activity is 75% of wild-type activity
W140T
-
DNA hydrolysis activity is 75% of wild-type activity
W140V
-
DNA hydrolysis activity is 75% of wild-type activity
W140Y
Y54C/I139C
-
production by site-directed mutagenesis, the oxidized form assumes a more compact denatured structure under acidic conditions than the wild type, the kinetic measurements reveal that the refolding reactions of both the reduced and oxidized forms of mutant are similar to those of the wild type protein
Y54C/I139C/DELTA140-149
-
production by site-directed mutagenesis, under physiological conditions, the reduced form appears to assume a denatured structure, in contrast, the oxidized form forms a native-like structure
Y91A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy)
Y91F
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
Y93A
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
Y93F
-
single point mutation in D21N/T33V/T41I/S59A/P117G/A128A, designed to examine short-range (up to 6.4 Angstrom) Coulomb and hydrogen bonding effects on His121, pKa values of histidines (His8, His46, His121, His124) are obtained by analysis of the pH titration monitored through the 1 H chemical shifts of the C(epsilon) H resonance of each histidine (NMR spectroscopy) , determination of tautomeric states of His121 and His124, pH near pI
Y93W/W140A
-
the mutant shows reduced activity with higher Michaelis-Menten constants, Km, and lower maximum reaction rate compared to the wild-type enzyme, the mutant also shows a more rapid loss of secondary and tertiary structure by Gdn-HCl unfolding than the wild-type enzyme
additional information