Q9LAI1 (T2BLA_BACSQ) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Type-2 restriction enzyme BslI subunit alpha
Short name=R.BslIalpha
EC=3.1.21.4

Alternative name(s):
Endonuclease BslI subunit alpha
Type II restriction enzyme BslI subunit alpha
Type IIT restriction enzyme BslI subunit alpha

Gene names

Name:

bslIRalpha

Organism

Bacillus sp. (strain NEB-606)

Taxonomic identifier

114630 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Firmicutes › Bacilli › Bacillales › Bacillaceae › Bacillus

Protein attributes

Sequence length	225 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Recognizes the double-stranded sequence 5'-CCN7GG-3' and cleaves after N-7.
Catalytic activity	Endonucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates.
Cofactor	Binds 2 zinc ions per subunit.
Subunit structure	Heterotetramer of two alpha and two beta subunits. The alpha subunit is believed to be responsible for DNA recognition, while the beta subunit is thought to mediate cleavage.
Domain	One of the zinc-binding motifs may participate in protein-DNA interactions and the other might mediate protein-protein interactions.
Biotechnological use	Could be used to screen carcinogenic mutations in a restriction endonuclease-mediated selective PCR (or REMS-PCR) assay. In particular, could be used to detect the vast majority of mutations that occur at codons 12 or 13 of the Ras oncogenes that encode glycine (codons GGN) at those positions.
Biophysicochemical properties	Temperature dependence: Is one of the thermostable restriction enzymes that remain active after 20 to 30 cycles of thermal PCR cycling.

Ontologies

Keywords
Biological process	Restriction system
Domain	Repeat Zinc-finger
Ligand	Metal-binding Zinc
Molecular function	Endonuclease Hydrolase Nuclease
Technical term	Direct protein sequencing
Gene Ontology (GO)
Biological_process	DNA restriction-modification system Inferred from electronic annotation. Source: UniProtKB-KW nucleic acid phosphodiester bond hydrolysis Inferred from electronic annotation. Source: GOC
Molecular_function	Type II site-specific deoxyribonuclease activity Inferred from electronic annotation. Source: EC metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 225	225	Type-2 restriction enzyme BslI subunit alpha		PRO_0000077283
Regions
	Zinc finger	36 – 53	18	C4-type 1
	Zinc finger	63 – 84	22	C4-type 2
Experimental info
	Mutagenesis	36	1	C → A: 10% of wild-type activity. Ref.2
	Mutagenesis	39	1	C → A: 10% of wild-type activity. Ref.2
	Mutagenesis	50	1	C → A: Loss of activity. Ref.2
	Mutagenesis	53	1	C → A: 10% of wild-type activity. Ref.1 Ref.2
	Mutagenesis	53	1	C → R: Loss of activity. Ref.1 Ref.2
	Mutagenesis	63	1	C → A: Loss of activity. Ref.2
	Mutagenesis	66	1	C → A: 50% of wild-type activity. Ref.2
	Mutagenesis	79	1	C → A: 50% of wild-type activity. Ref.2
	Mutagenesis	84	1	C → A: 50% of wild-type activity. Ref.2

Sequences

	Sequence		Length	Mass (Da)
	Q9LAI1 [UniParc]. Last modified October 1, 2000. Version 1. Checksum: 4969BF02325E12B2	FASTA	225	25,605
	10 20 30 40 50 60 MERQLKSIAY AFVANDIDVY IPDGESNCIV VTKLVCKDCG QYWHTSLSEC YFCGTLNFYL 70 80 90 100 110 120 YECNSCGKKY SLTSSSKSCD TDGCNGKLIK RCSNPECISR TNEEIQRATD EQGGVFDLNS 130 140 150 160 170 180 SFNVSLNHCV TCGSKENYYK TYRIYSYRTE VEPNIEALRE FANNNKLNSD EDVIIIKHLV 190 200 210 220 DNVIHYGYIP YSKLDETTEI TTTFSRFSDL VSELFPVNVP PNVTE

References

[1]	"Cloning, expression, and purification of a thermostable nonhomodimeric restriction enzyme, BslI." Hsieh P.-C., Xiao J.-P., O'Loane D., Xu S.-Y. J. Bacteriol. 182:949-955(2000) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF N-TERMINUS, CHARACTERIZATION, MUTAGENESIS OF CYS-53.
[2]	"Glucocorticoid receptor-like Zn(Cys)4 motifs in BslI restriction endonuclease." Scheuring Vanamee E., Hsieh P.-C., Zhu Z., Yates D., Garman E., Xu S.-Y., Aggarwal A.K. J. Mol. Biol. 334:595-603(2003) [PubMed] [Europe PMC] [Abstract] Cited for: ZINC-BINDING, ABSORPTION SPECTROSCOPY, MUTAGENESIS OF CYSTEINE RESIDUES.

Cross-references

Sequence databases
EMBL GenBank DDBJ	AF135191 Genomic DNA. Translation: AAF32530.1.
3D structure databases
ModBase	Search...
Protein family/group databases
REBASE	386. BslI.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Family and domain databases
ProtoNet	Search...

Entry information

Entry name

T2BLA_BACSQ

Accession

Primary (citable) accession number: Q9LAI1

Entry history

Integrated into UniProtKB/Swiss-Prot:	February 16, 2004
Last sequence update:	October 1, 2000
Last modified:	April 3, 2013
This is version 34 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program