T2E1_ECOLX
ID T2E1_ECOLX Reviewed; 277 AA.
AC P00642;
DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 2.
DT 03-AUG-2022, entry version 137.
DE RecName: Full=Type II restriction enzyme EcoRI {ECO:0000303|PubMed:12654995};
DE Short=R.EcoRI;
DE EC=3.1.21.4 {ECO:0000269|PubMed:3024321};
DE AltName: Full=Endonuclease EcoRI;
DE AltName: Full=Type-2 restriction enzyme EcoRI;
GN Name=ecoRIR;
OS Escherichia coli.
OG Plasmid pMB1, and Plasmid pMB4.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC PLASMID=pMB1;
RX PubMed=6257703; DOI=10.1016/s0021-9258(19)69752-8;
RA Greene P.J., Gupta M., Boyer H.W., Brown W.E., Rosenberg J.M.;
RT "Sequence analysis of the DNA encoding the Eco RI endonuclease and
RT methylase.";
RL J. Biol. Chem. 256:2143-2153(1981).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=C; PLASMID=pMB4;
RX PubMed=6257701; DOI=10.1016/s0021-9258(19)69750-4;
RA Newman A.K., Rubin R.A., Kim S.-H., Modrich P.;
RT "DNA sequences of structural genes for Eco RI DNA restriction and
RT modification enzymes.";
RL J. Biol. Chem. 256:2131-2139(1981).
RN [3]
RP PROTEIN SEQUENCE OF 2-13, AND PROTEIN SEQUENCE OF C-TERMINUS.
RX PubMed=6257702; DOI=10.1016/s0021-9258(19)69751-6;
RA Rubin R.A., Modrich P., Vanaman T.C.;
RT "Partial NH2- and COOH-terminal sequence analyses of Eco RI DNA restriction
RT and modification enzymes.";
RL J. Biol. Chem. 256:2140-2142(1981).
RN [4]
RP FAMILY.
RX PubMed=2843805; DOI=10.1093/nar/16.16.7901;
RA Aiken C., Gumport R.I.;
RT "Restriction endonuclease RsrI from Rhodobacter sphaeroides, an
RT isoschizomer of EcoRI: purification and properties.";
RL Nucleic Acids Res. 16:7901-7916(1988).
RN [5]
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).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (3.0 ANGSTROMS), AND SUBUNIT.
RX PubMed=3024321; DOI=10.1126/science.3024321;
RA McClarin J.A., Frederick C.A., Wang B.-C., Greene P., Boyer H.W.,
RA Grable J., Rosenberg J.M.;
RT "Structure of the DNA-Eco RI endonuclease recognition complex at 3-A
RT resolution.";
RL Science 234:1526-1541(1986).
RN [7] {ECO:0007744|PDB:1ERI}
RP X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS).
RX PubMed=2399465; DOI=10.1126/science.2399465;
RA Kim Y., Grable J.C., Love R., Greene P.J., Rosenberg J.M.;
RT "Refinement of Eco RI endonuclease crystal structure: a revised protein
RT chain tracing.";
RL Science 249:1307-1309(1990).
RN [8]
RP REVIEW.
RA Rosenberg J.M.;
RT "Structure and function of restriction endonucleases.";
RL Curr. Opin. Struct. Biol. 1:104-113(1991).
RN [9]
RP MUTAGENESIS OF GLU-144.
RX PubMed=2254311; DOI=10.1016/s0021-9258(18)45770-5;
RA Hager P.W., Reich N.O., Day J.P., Coche T.G., Boyer H.W., Rosenberg J.M.,
RA Greene P.J.;
RT "Probing the role of glutamic acid 144 in the EcoRI endonuclease using
RT aspartic acid and glutamine replacements.";
RL J. Biol. Chem. 265:21520-21526(1990).
CC -!- FUNCTION: A P subtype restriction enzyme that recognizes the double-
CC stranded sequence 5'-GAATTC-3' and cleaves after G-1.
CC {ECO:0000269|PubMed:3024321, 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:3024321};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Note=Binds 2 magnesium ions per subunit.;
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:3024321}.
CC -!- SIMILARITY: Belongs to the EcoRI type II restriction endonuclease
CC family. {ECO:0000303|PubMed:2843805}.
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DR EMBL; J01675; AAA26371.1; -; Genomic_DNA.
DR PIR; B92308; NDECP4.
DR RefSeq; WP_001565219.1; NZ_SQMQ01000003.1.
DR PDB; 1CKQ; X-ray; 1.85 A; A=2-277.
DR PDB; 1CL8; X-ray; 1.80 A; A=2-277.
DR PDB; 1ERI; X-ray; 2.50 A; A=2-277.
DR PDB; 1QC9; X-ray; 3.00 A; A/B/C=2-277.
DR PDB; 1QPS; X-ray; 2.50 A; A=17-277.
DR PDB; 1QRH; X-ray; 2.50 A; A=17-277.
DR PDB; 1QRI; X-ray; 2.60 A; A=17-277.
DR PDB; 2OXV; X-ray; 1.95 A; A=1-277.
DR PDBsum; 1CKQ; -.
DR PDBsum; 1CL8; -.
DR PDBsum; 1ERI; -.
DR PDBsum; 1QC9; -.
DR PDBsum; 1QPS; -.
DR PDBsum; 1QRH; -.
DR PDBsum; 1QRI; -.
DR PDBsum; 2OXV; -.
DR AlphaFoldDB; P00642; -.
DR SMR; P00642; -.
DR DIP; DIP-16997N; -.
DR BindingDB; P00642; -.
DR ChEMBL; CHEMBL5729; -.
DR REBASE; 993; EcoRI.
DR PATRIC; fig|562.8010.peg.1144; -.
DR BRENDA; 3.1.21.4; 2026.
DR EvolutionaryTrace; P00642; -.
DR PRO; PR:P00642; -.
DR GO; GO:0003677; F:DNA binding; IEA:InterPro.
DR GO; GO:0000287; F:magnesium ion binding; IEA:InterPro.
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.
DR Gene3D; 3.40.580.10; -; 1.
DR InterPro; IPR011335; Restrct_endonuc-II-like.
DR InterPro; IPR004221; Restrct_endonuc_II_EcoRI.
DR InterPro; IPR011336; Restrct_endonuc_II_EcoRI/MunI.
DR InterPro; IPR018131; Restrct_endonuc_II_EcoRI_Pbac.
DR Pfam; PF02963; EcoRI; 1.
DR PIRSF; PIRSF001002; Restrict_endonuc_II_EcoRI; 1.
DR SUPFAM; SSF52980; SSF52980; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Endonuclease; Hydrolase;
KW Magnesium; Metal-binding; Nuclease; Plasmid; Restriction system.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:6257702"
FT CHAIN 2..277
FT /note="Type II restriction enzyme EcoRI"
FT /id="PRO_0000077303"
FT ACT_SITE 91
FT ACT_SITE 111
FT ACT_SITE 113
FT BINDING 91
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT BINDING 91
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT BINDING 111
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT MUTAGEN 144
FT /note="E->D: Only nicks double strand DNA."
FT /evidence="ECO:0000269|PubMed:2254311"
FT MUTAGEN 144
FT /note="E->Q: Inactivates the enzyme."
FT /evidence="ECO:0000269|PubMed:2254311"
FT HELIX 21..31
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 35..49
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 55..59
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 63..73
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 94..98
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 104..114
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 118..123
FT /evidence="ECO:0007829|PDB:1CL8"
FT TURN 129..132
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 134..136
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 142..145
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 146..156
FT /evidence="ECO:0007829|PDB:1CL8"
FT TURN 157..159
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 160..162
FT /evidence="ECO:0007829|PDB:1CKQ"
FT STRAND 165..171
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 172..174
FT /evidence="ECO:0007829|PDB:1QPS"
FT STRAND 179..182
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 184..186
FT /evidence="ECO:0007829|PDB:1ERI"
FT STRAND 188..191
FT /evidence="ECO:0007829|PDB:1CL8"
FT TURN 196..198
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 201..203
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 205..208
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 213..215
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 221..223
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 225..227
FT /evidence="ECO:0007829|PDB:1CKQ"
FT STRAND 230..232
FT /evidence="ECO:0007829|PDB:1CL8"
FT STRAND 236..239
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 248..265
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 267..269
FT /evidence="ECO:0007829|PDB:1CL8"
FT HELIX 271..275
FT /evidence="ECO:0007829|PDB:1CL8"
SQ SEQUENCE 277 AA; 31059 MW; 83EDC82868261B4B CRC64;
MSNKKQSNRL TEQHKLSQGV IGIFGDYAKA HDLAVGEVSK LVKKALSNEY PQLSFRYRDS
IKKTEINEAL KKIDPDLGGT LFVSNSSIKP DGGIVEVKDD YGEWRVVLVA EAKHQGKDII
NIRNGLLVGK RGDQDLMAAG NAIERSHKNI SEIANFMLSE SHFPYVLFLE GSNFLTENIS
ITRPDGRVVN LEYNSGILNR LDRLTAANYG MPINSNLCIN KFVNHKDKSI MLQAASIYTQ
GDGREWDSKI MFEIMFDIST TSLRVLGRDL FEQLTSK
