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 04-NOV-2026, entry version 145. 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; Pseudomonadati; Pseudomonadota; Gammaproteobacteria; OC Enterobacterales; 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, 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. 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}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; J01675; AAA26371.1; -; Genomic_DNA. DR PIR; B92308; NDECP4. DR RefSeq; WP_001565219.1; NZ_BKCB01000112.1. 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 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 CDD; cd00943; EcoRI-like; 1. DR Gene3D; 3.40.580.10; Eco RI Endonuclease, subunit A; 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; Restriction endonuclease-like; 1. 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; EXP:UniProtKB. DR GO; GO:0009307; P:DNA restriction-modification system; IEA:InterPro. DR AlphaFoldDB; P00642; -. DR MDposit; P00642; -. 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 244..247 FT /evidence="ECO:0007829|PDB:1QRH" 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 //