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3.1.21.4: type II site-specific deoxyribonuclease

This is an abbreviated version!
For detailed information about type II site-specific deoxyribonuclease, go to the full flat file.

Word Map on EC 3.1.21.4

Reaction

endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates =

Synonyms

AatII, AcuI, acuIRM, AfeI, AflII, AflIII, aflIIR, AgeI, AhaIII, AlfI, AloI, AluI, AlwNI, Apa BI, Apa CI, Apa DI, ApaI, ApaLI, apaLIR, AsuI, AsuII, AvaI, AvaII, AvaIII, AvrII, BaeI, BamHI, bamHIR, BbeI, BbvCI, BbvI, BbvII, bbvIR, BcefI, BcgI, BclI, Bfi2411I, BgII, BglI, BglII, bglIIR, BinI, BmrI, BplI, BpmI, bpmIR, BpuEI, BpuIOI, BsaAI, BsaBI, BsaWI, BsaXI, Bse634I, BsePI, BseRI, BseRI endonuclease, BsgI, BsmAI, BsmBI, BsmI, BsoBI, Bsp CI, BspD6I, BspGI, BspHI, BspI, BspMI, BSPMII, BspRI, BsrI, BssSI, BstEII, BstF5I, BstII, BstXI, BstYI, Bsu2413I, BtsI, CauII, Cfr101, Cfr10I, CfrBI, CfrI, CjeI, ClaI, CspCI, CstMI, CviAI, CviJI, DdeI, DNA restriction endonuclease, DNA restriction enzyme, DpnI, DpnII, DraII, DraIII, DrdI, DrdII, DsaI, EC 3.1.23, EC 3.1.24, EciI, Eco1524I, Eco311, Eco31I, Eco47III, Eco571, Eco57I, eco57IR, EcoO109I, EcoP15I, ecoP15Ires, EcoRI, EcoRII, ecoRIR, EcoRV, EcoVIII, EgeI, EheI, Endonuclease AbrI, Endonuclease AccI, Endonuclease AgeI, Endonuclease ApaLI, Endonuclease AvaI, Endonuclease BamHI, Endonuclease BanI, Endonuclease BglI, Endonuclease BglII, Endonuclease BsoBI, Endonuclease Bsp6I, Endonuclease BstVI, Endonuclease BsuBI, Endonuclease BsuFI, Endonuclease BsuRI, Endonuclease CeqI, Endonuclease Cfr10I, Endonuclease Cfr9I, Endonuclease CfrBI, Endonuclease CviAII, Endonuclease CviJI, Endonuclease DdeI, Endonuclease DpnI, Endonuclease DpnII, Endonuclease Eco47I, Endonuclease Eco47II, Endonuclease EcoRI, Endonuclease EcoRII, Endonuclease EcoRV, Endonuclease FokI, Endonuclease HaeII, Endonuclease HaeIII, Endonuclease HgAI, Endonuclease HgiBI, Endonuclease HgiCI, Endonuclease HgiCII, Endonuclease HgiDI, Endonuclease HgiEI, Endonuclease HgiGI, Endonuclease HhaII, Endonuclease HincII, Endonuclease HindII, Endonuclease HindIII, Endonuclease HindVP, Endonuclease HinfI, Endonuclease HpaI, Endonuclease HpaII, Endonuclease HphI, Endonuclease KpnI, Endonuclease LlaDCHI, Endonuclease MamI, Endonuclease MboI, Endonuclease MboII, Endonuclease MjaI, Endonuclease MjaII, Endonuclease MjaIII, Endonuclease MjaIV, Endonuclease MjaV, Endonuclease MjaVIP, Endonuclease MspI, Endonuclease MthTI, Endonuclease MthZI, Endonuclease MunI, Endonuclease MwoI, Endonuclease NaeI, Endonuclease NgoBI, Endonuclease NgoBV, Endonuclease NgoFVII, Endonuclease NgoMIV, Endonuclease NgoPII, Endonuclease NlaIII, Endonuclease NlaIV, Endonuclease NmeDIP, Endonuclease NspV, Endonuclease PaeR7I, Endonuclease PstI, Endonuclease PvuI, Endonuclease PvuII, Endonuclease RsrI, Endonuclease SacI, Endonuclease SalI, Endonuclease Sau3AI, Endonuclease Sau96I, Endonuclease ScaI, Endonuclease ScrFI, Endonuclease SfiI, Endonuclease SinI, Endonuclease SmaI, Endonuclease SsoII, Endonuclease StsI, Endonuclease TaqI, Endonuclease TthHB8I, Endonuclease XamI, Endonuclease XcyI, endoR*Bsp, Esp3I, EspI, FauI, FinI, Fnu4HI, FnuSII, FokI, fokIR, FseI, GdiII, GGCC-specific restriction endonuclease, GsuI, HaeI, HaeII, HaeIII, HaeVI, HgaI, HgiAI, HgiCI, HgiEI, HgiEII, HgiJII, HhaI, HhaII, HincII, HindII, HindIII, hindIIIR, HinfI, HinP1I, HpaI, HpaII, HphI, Hpy188I, Hpy8I, Hpy99II, Hpy99IV, Hpy99VIIIP, HpyAXII, HpyF17I, KasI, KpnI, KpnIR, Ksp6321, LlaDI, LlaII, LmoJ2, LmoJ3, MaeI, MaeII, MaeIII, MboI, MboII, McaTI, McrI, MfeI, MjaIV, MjaV, MluI, Mly113I, MlyI, MmeI, mmeIRM, MmoSTI, MnlI, More, MseI, MspA1I, MspI, MstI, MunI, Mva1269I, Mva1269I restriction endonuclease, MvaI, MwoI, NaeI, NarI, NcoI, ncoIR, NdeI, NgoMIV, NheI, NlaIII, NlaIV, NmeDI, NmeDIP, NotI, NruI, NspBII, NspV, Nt.BspD6I, nuclease, deoxyribonucleic restriction endo-, nuclease, restriction endodeoxyribo-, PabI, PflMI, PhoI, PleI, PmaCI, PPII, PpuMI, PsasM2I, PshAI, PspGI, PstI, pstIR, PvuI, PvuII, R.AbrI, R.AccI, R.AgeI, R.ApaLI, R.AvaI, R.BamHI, R.BanI, R.BglI, R.BglII, R.BsoBI, R.Bsp6I, R.BstVI, R.BsuBI, R.BsuFI, R.BsuRI, R.CeqI, R.Cfr10I, R.Cfr9I, R.CfrBI, R.CviAII, R.CviJI, R.DdeI, R.DpnI, R.DpnII, R.Eco47I, R.Eco47II, R.EcoRI, R.EcoRII, R.EcoRV, R.FokI, R.HaeII, R.HaeIII, R.HgAI, R.HgiBI, R.HgiCI, R.HgiCII, R.HgiDI, R.HgiEI, R.HgiGI, R.HhaII, R.HincII, R.HindII, R.HindIII, R.HindVP, R.HinfI, R.HpaI, R.HpaII, R.HphI, R.KpnI, R.LlaDCHI, R.MamI, R.MboI, R.MboII, R.MjaI, R.MjaII, R.MjaIII, R.MjaIV, R.MjaV, R.MjaVIP, R.MspI, R.MthTI, R.MthZI, R.MunI, R.MwoI, R.NaeI, R.NgoBI, R.NgoBV, R.NgoFVII, R.NgoI, R.NgoMIV, R.NgoPII, R.NgoV, R.NgoVII, R.NlaIII, R.NlaIV, R.NmeDIP, R.NspV, R.PaeR7I, R.PstI, R.PvuI, R.PvuII, R.RsrI, R.SacI, R.SalI, R.Sau3AI, R.Sau96I, R.ScaI, R.ScrFI, R.SfiI, R.SinI, R.SmaI, R.SsoII, R.StsI, R.TaqI, R.TthHB8I, R.XamI, R.XcyI, REase, restriction endodeoxyribonuclease, restriction endonuclease, restriction endonuclease EcoRV, restriction endonuclease SuaI, restriction enzyme, RleAI, RsaI, SacI, SacII, SacIR, SalI, Sau3AI, sau96I, SauI, ScaI, ScrFI, SduI, SecI, SfaNI, SfeI, SfiI, SfoI, SgrAI, Sho27844P, site-specific endonuclease BME142I, site-specific endonuclease Bme216I, site-specific endonuclease BmeI, site-specific type II endonuclease, SmaI, SmuI, SnaBI, SnaI, SpeI, SphI, SpiI, ss.BspD6I, Sse9I, SspI, StuI, StyI, SuaI, SuiI, TaqI, TaqII, TfiI, Tsp451, TspEI, TstI, Tth111I, Tth111II, type II ENase, type II REase, type II restriction endonuclease, type II restriction endonuclease EcoO109I, type II restriction enonuclease, type II restriction enzyme, type II restriction-modification protein, type II restriction-modification system, type II RM system, type IIB restriction endonuclease, type IIE restriction endonuclease, type IIF restriction endonuclease, type IIP REase, type IIP restriction endonuclease, Type IIS REase, type IIS restriction endonuclease, type IIS restriction enzyme, type IIT restriction endonuclease BbvCI, typeIIS restriction endonuclease, Uba1105I, Uba1108I, UbaLAI, UpaP162, UPV_230, VspI, XbaI, XcmI, XhoI, XhoII, XmaIII, XmnI

ECTree

     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.21 Endodeoxyribonucleases producing 5'-phosphomonoesters
                3.1.21.4 type II site-specific deoxyribonuclease

Engineering

Engineering on EC 3.1.21.4 - type II site-specific deoxyribonuclease

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D140A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
D140A/E167A/K169A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
D144A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit
D146A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit
D146A/E177A
Q5D6Y5; Q5D6Y4
slight detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit
D146A/E177A/K179A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit
D149A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit
D226A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit
E139A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit
E147A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit
E155A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, low activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
E163A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit
E165A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit
E167A
Q5D6Y5; Q5D6Y4
slight activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
E167A/K169A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
E167G
Q5D6Y5; Q5D6Y4
slight activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
E177A
Q5D6Y5; Q5D6Y4
slight activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit
E177G
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit
E201A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit
E211A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking and no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit
E218A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, no activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
E227A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking and no detectable activity in ssDNA nicking, mutant R2 subunit/wild-type R1 subunit
E95A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA and in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
K169A
Q5D6Y5; Q5D6Y4
low activity in dsDNA nicking, mutant R1 subunit/wild-type R2 subunit
K179A
Q5D6Y5; Q5D6Y4
no detectable activity in dsDNA nicking, mutant R2 subunit/wild-type R1 subunit
N138A
Q5D6Y5; Q5D6Y4
no detectable activity in ssDNA nicking, mutant R1 subunit/wild-type R2 subunit
D134N/K190N/E204V
-
inactive
E204Q
-
the mutant possesses 0.004% of wild type activity
E71A/K190T/E204I
-
inactive
E71A/K190T/E204V
-
inactive
E71A/S188A/K190T/E204Q
-
inactive
E71N/D254G
-
inactive
E71Q
-
the mutant possesses 10% of wild type activity
S188A
-
the mutant possesses 4% of wild type activity
S188C
-
the mutant possesses 0.5% of wild type activity
D299A
-
no catalytic activity, mutant like wild-type remains in dimeric form
D329A
-
less than 0.01% residual activity, mutant like wild-type remains in dimeric form
E271A
-
no catalytic activity, mutant like wild-type remains in dimeric form
E337A
-
less than 0.01% residual activity, mutant like wild-type remains in dimeric form
K263A
-
inactive mutant with truncated EcoRII N-domain
K324A
-
no catalytic activity, mutant like wild-type remains in dimeric form
K328A
-
less than 0.01% residual activity, mutant like wild-type remains in dimeric form
L80P
the mutant enzyme shows decreased DNA methyltransferase activity at a higher temperature in vivo and in vitro than the wild type enzyme, the activity of the L80P mutant is completely lost at a high temperature
M357P
-
mutation converts DNA methyltransferase to a type II endonuclease with 5'-CAGCAG-3' restriction site that cleaves only supercoiled DNA but does not act on nicked or linearized DNA
R330A
R88A
-
crystallization data, autoinhibition/activation mechanism
L80P
-
the mutant enzyme shows decreased DNA methyltransferase activity at a higher temperature in vivo and in vitro than the wild type enzyme, the activity of the L80P mutant is completely lost at a high temperature
-
DELTA362-465
-
nicking variant of BsaI, nicking occurs on top strand
DELTA440-544
-
nicking variant of BsaI, nicking occurs on bottom strand
DELTA446-544
-
nicking variant of BsaI
K150R/R236G
-
nicking variant of BsaI, nicking occurs on top strand
N349Y
-
nicking variant of BsmAI, nicking occurs on top strand
N415D/R416G
-
variant of BsmAI, both nicking activity and dsDNA cleavage
N441D
-
variant of BsaI, cleavage of dsDNA
N441D/R442G
-
nicking variant of BsaI, nicking occurs on bottom strand
R221D
-
nicking variant of BsmAI, nicking on top strand
R233D
-
nicking variant of BsmBI, nicking on top strand, about 5-10% cleavage of dsDNA
R236D
-
nicking variant of BsaI, nicking on top strand, no cleavage of dsDNA
R236G
-
nicking variant of BsaI, 5-10% cleavage of dsDNA
R386S
-
nicking variant of BsaI, nicking occurs on top strand
R438D
-
nicking variant of BsmBI, nicking on bottom strand
R438G
-
nicking variant of BsmBI
R442G
-
nicking variant of BsaI, 5% cleavage of dsDNA
S128L/R236G
-
nicking variant of BsaI, nicking occurs on top strand
W8C/G207E
-
nicking variant of BsaI, nicking occurs on top strand
Q138F
mutation results in a change in the sequence specificity at the center two base pairs of the cognate recognition site. Alteration in preference of HicII for cutting, but not binding, the three cognate sites differening in the center two base pairs.The Q138F HincII/DNA crystal structures show conformational changes in the protein, bound DNA, and at the protein-DNA interface
D317A
-
84% of DNA binding compared to wild-type
D321A
-
119% of DNA binding compared to wild-type
D328A
-
76% of DNA binding compared to wild-type
D329A
-
123% of DNA binding compared to wild-type
H291A
-
96% of DNA binding compared to wild-type
H368A
-
129% of DNA binding compared to wild-type
N308A
-
104% of DNA binding compared to wild-type
D148A
-
only traces of DNA cleavage activity when used in large excess
D148G
H149L
Q115E
-
deprotonated mutant EcoRI is defective in DNA binding at neutral pH
Q175E
-
only traces of DNA cleavage activity when used in large excess
R119A
-
subunit BtsIB mutant shows a different digestion pattern from the wild type BtsI. The mutant BtsIB(R119A) acts as a different restriction enzyme with a previously unreported recognition sequence CAGTG(2/0), which is named as BtsI-1
E64A
-
monomer crystallizes in space group C2221, two molecules per asymmetric unit
additional information