T2BLA_BACSQ
ID T2BLA_BACSQ Reviewed; 225 AA.
AC Q9LAI1;
DT 16-FEB-2004, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2000, sequence version 1.
DT 25-MAY-2022, entry version 49.
DE RecName: Full=Type II restriction enzyme BslI subunit alpha {ECO:0000303|PubMed:12654995};
DE Short=R.BslIalpha;
DE EC=3.1.21.4 {ECO:0000269|PubMed:10648519};
DE AltName: Full=Endonuclease BslI subunit alpha;
DE AltName: Full=Type IIT restriction enzyme BslI subunit alpha;
DE AltName: Full=Type-2 restriction enzyme BslI subunit alpha;
GN Name=bslIRalpha {ECO:0000303|PubMed:10648519};
OS Bacillus sp. (strain NEB-606).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=114630;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF N-TERMINUS,
RP FUNCTION, SUBUNIT, AND MUTAGENESIS OF CYS-53.
RX PubMed=10648519; DOI=10.1128/jb.182.4.949-955.2000;
RA Hsieh P.-C., Xiao J.-P., O'Loane D., Xu S.-Y.;
RT "Cloning, expression, and purification of a thermostable nonhomodimeric
RT restriction enzyme, BslI.";
RL J. Bacteriol. 182:949-955(2000).
RN [2]
RP ZINC-BINDING, ABSORPTION SPECTROSCOPY, AND MUTAGENESIS OF CYS-36; CYS-39;
RP CYS-50; CYS-53; CYS-63; CYS-66; CYS-79 AND CYS-84.
RX PubMed=14623197; DOI=10.1016/j.jmb.2003.09.043;
RA Scheuring Vanamee E., Hsieh P.-C., Zhu Z., Yates D., Garman E., Xu S.-Y.,
RA Aggarwal A.K.;
RT "Glucocorticoid receptor-like Zn(Cys)4 motifs in BslI restriction
RT endonuclease.";
RL J. Mol. Biol. 334:595-603(2003).
RN [3]
RP NOMENCLATURE, AND SUBTYPE.
RX PubMed=12654995; DOI=10.1093/nar/gkg274;
RA Roberts R.J., Belfort M., Bestor T., Bhagwat A.S., Bickle T.A.,
RA Bitinaite J., Blumenthal R.M., Degtyarev S.K., Dryden D.T., Dybvig K.,
RA Firman K., Gromova E.S., Gumport R.I., Halford S.E., Hattman S.,
RA Heitman J., Hornby D.P., Janulaitis A., Jeltsch A., Josephsen J., Kiss A.,
RA Klaenhammer T.R., Kobayashi I., Kong H., Krueger D.H., Lacks S.,
RA Marinus M.G., Miyahara M., Morgan R.D., Murray N.E., Nagaraja V.,
RA Piekarowicz A., Pingoud A., Raleigh E., Rao D.N., Reich N., Repin V.E.,
RA Selker E.U., Shaw P.C., Stein D.C., Stoddard B.L., Szybalski W.,
RA Trautner T.A., Van Etten J.L., Vitor J.M., Wilson G.G., Xu S.Y.;
RT "A nomenclature for restriction enzymes, DNA methyltransferases, homing
RT endonucleases and their genes.";
RL Nucleic Acids Res. 31:1805-1812(2003).
CC -!- FUNCTION: A P subtype restriction enzyme that recognizes the double-
CC stranded sequence 5'-CCN(7)GG-3' and cleaves after N-7.
CC {ECO:0000269|PubMed:10648519, ECO:0000303|PubMed:12654995}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Endonucleolytic cleavage of DNA to give specific double-
CC stranded fragments with terminal 5'-phosphates.; EC=3.1.21.4;
CC Evidence={ECO:0000269|PubMed:10648519};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Note=Binds 2 Zn(2+) ions per subunit. {ECO:0000269|PubMed:14623197};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Temperature dependence:
CC Is one of the thermostable restriction enzymes that remain active
CC after 20 to 30 cycles of thermal PCR cycling.;
CC -!- SUBUNIT: Heterotetramer of two alpha and two beta subunits. The alpha
CC subunit is believed to be responsible for DNA recognition, while the
CC beta subunit is thought to mediate cleavage.
CC {ECO:0000269|PubMed:10648519}.
CC -!- DOMAIN: One of the zinc-binding motifs may participate in protein-DNA
CC interactions and the other might mediate protein-protein interactions.
CC -!- BIOTECHNOLOGY: Could be used to screen carcinogenic mutations in a
CC restriction endonuclease-mediated selective PCR (or REMS-PCR) assay. In
CC particular, could be used to detect the vast majority of mutations that
CC occur at codons 12 or 13 of the Ras oncogenes that encode glycine
CC (codons GGN) at those positions.
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DR EMBL; AF135191; AAF32530.1; -; Genomic_DNA.
DR AlphaFoldDB; Q9LAI1; -.
DR REBASE; 386; BslI.
DR PRO; PR:Q9LAI1; -.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0009036; F:type II site-specific deoxyribonuclease activity; IEA:UniProtKB-EC.
DR GO; GO:0009307; P:DNA restriction-modification system; IEA:UniProtKB-KW.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Endonuclease; Hydrolase; Metal-binding;
KW Nuclease; Repeat; Restriction system; Zinc; Zinc-finger.
FT CHAIN 1..225
FT /note="Type II restriction enzyme BslI subunit alpha"
FT /id="PRO_0000077283"
FT ZN_FING 36..53
FT /note="C4-type 1"
FT ZN_FING 63..84
FT /note="C4-type 2"
FT MUTAGEN 36
FT /note="C->A: 10% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:14623197"
FT MUTAGEN 39
FT /note="C->A: 10% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:14623197"
FT MUTAGEN 50
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:14623197"
FT MUTAGEN 53
FT /note="C->A: 10% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:10648519"
FT MUTAGEN 53
FT /note="C->R: Loss of activity."
FT /evidence="ECO:0000269|PubMed:10648519"
FT MUTAGEN 63
FT /note="C->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:14623197"
FT MUTAGEN 66
FT /note="C->A: 50% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:14623197"
FT MUTAGEN 79
FT /note="C->A: 50% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:14623197"
FT MUTAGEN 84
FT /note="C->A: 50% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:14623197"
SQ SEQUENCE 225 AA; 25605 MW; 4969BF02325E12B2 CRC64;
MERQLKSIAY AFVANDIDVY IPDGESNCIV VTKLVCKDCG QYWHTSLSEC YFCGTLNFYL
YECNSCGKKY SLTSSSKSCD TDGCNGKLIK RCSNPECISR TNEEIQRATD EQGGVFDLNS
SFNVSLNHCV TCGSKENYYK TYRIYSYRTE VEPNIEALRE FANNNKLNSD EDVIIIKHLV
DNVIHYGYIP YSKLDETTEI TTTFSRFSDL VSELFPVNVP PNVTE
