BLAB_AERHY
ID BLAB_AERHY Reviewed; 254 AA.
AC P26918;
DT 01-AUG-1992, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1992, sequence version 1.
DT 03-AUG-2022, entry version 115.
DE RecName: Full=Metallo-beta-lactamase type 2 {ECO:0000305};
DE EC=3.5.2.6 {ECO:0000269|PubMed:15588826, ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:2327760};
DE AltName: Full=B2 metallo-beta-lactamase {ECO:0000305};
DE AltName: Full=Beta-lactamase II {ECO:0000303|PubMed:1856163};
DE AltName: Full=Carbapenem-hydrolyzing metallo-beta-lactamase {ECO:0000303|PubMed:1856163};
DE AltName: Full=Metallo-beta-lactamase type II {ECO:0000303|PubMed:1856163};
DE Flags: Precursor;
GN Name=cphA {ECO:0000303|PubMed:1856163};
OS Aeromonas hydrophila.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Aeromonadales;
OC Aeromonadaceae; Aeromonas.
OX NCBI_TaxID=644;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, ACTIVITY REGULATION, SUBSTRATE
RP SPECIFICITY, AND SUBUNIT.
RC STRAIN=AE036;
RX PubMed=1856163; DOI=10.1128/jb.173.15.4611-4617.1991;
RA Massidda O., Rossolini G.M., Satta G.;
RT "The Aeromonas hydrophila cphA gene: molecular heterogeneity among class B
RT metallo-beta-lactamases.";
RL J. Bacteriol. 173:4611-4617(1991).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, INDUCTION, SUBUNIT, AND SUBSTRATE SPECIFICITY.
RX PubMed=2327760; DOI=10.1128/aac.34.1.44;
RA Iaconis J.P., Sanders C.C.;
RT "Purification and characterization of inducible beta-lactamases in
RT Aeromonas spp.";
RL Antimicrob. Agents Chemother. 34:44-51(1990).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF 28-254 OF WILD-TYPE AND MUTANT
RP GLY-192 IN COMPLEX WITH SUBSTRATE ANALOGS AND ZINC IONS, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ASN-192,
RP COFACTOR, SUBUNIT, AND REACTION MECHANISM.
RX PubMed=15588826; DOI=10.1016/j.jmb.2004.10.070;
RA Garau G., Bebrone C., Anne C., Galleni M., Frere J.M., Dideberg O.;
RT "A metallo-beta-lactamase enzyme in action: crystal structures of the
RT monozinc carbapenemase CphA and its complex with biapenem.";
RL J. Mol. Biol. 345:785-795(2005).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.86 ANGSTROMS) OF 28-254 IN COMPLEX WITH SUBSTRATE
RP ANALOG AND ZINC IONS, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, ACTIVITY REGULATION, MUTAGENESIS OF ASN-93 AND ASN-192, AND
RP COFACTOR.
RX PubMed=17307979; DOI=10.1128/aac.00866-06;
RA Horsfall L.E., Garau G., Lienard B.M., Dideberg O., Schofield C.J.,
RA Frere J.M., Galleni M.;
RT "Competitive inhibitors of the CphA metallo-beta-lactamase from Aeromonas
RT hydrophila.";
RL Antimicrob. Agents Chemother. 51:2136-2142(2007).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.66 ANGSTROMS) OF 28-254 IN COMPLEX WITH SUBSTRATE
RP ANALOG AND ZINC IONS, AND COFACTOR.
RX PubMed=18563261; DOI=10.1039/b802311e;
RA Lienard B.M., Garau G., Horsfall L., Karsisiotis A.I., Damblon C.,
RA Lassaux P., Papamicael C., Roberts G.C., Galleni M., Dideberg O.,
RA Frere J.M., Schofield C.J.;
RT "Structural basis for the broad-spectrum inhibition of metallo-beta-
RT lactamases by thiols.";
RL Org. Biomol. Chem. 6:2282-2294(2008).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) OF 28-254 OF WILD-TYPE AND MUTANT
RP GLY-192 IN COMPLEX WITH SUBSTRATE ANALOGS AND ZINC IONS, FUNCTION,
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ASN-192,
RP ACTIVITY REGULATION, AND COFACTOR.
RX PubMed=19651913; DOI=10.1128/aac.00288-09;
RA Bebrone C., Delbruck H., Kupper M.B., Schlomer P., Willmann C., Frere J.M.,
RA Fischer R., Galleni M., Hoffmann K.M.;
RT "The structure of the dizinc subclass B2 metallo-beta-lactamase CphA
RT reveals that the second inhibitory zinc ion binds in the histidine site.";
RL Antimicrob. Agents Chemother. 53:4464-4471(2009).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.41 ANGSTROMS) OF 28-254 IN COMPLEX WITH SUBSTRATE
RP ANALOGS AND ZINC IONS, ACTIVITY REGULATION, AND COFACTOR.
RX PubMed=20527888; DOI=10.1021/jm100213c;
RA Lassaux P., Hamel M., Gulea M., Delbruck H., Mercuri P.S., Horsfall L.,
RA Dehareng D., Kupper M., Frere J.M., Hoffmann K., Galleni M., Bebrone C.;
RT "Mercaptophosphonate compounds as broad-spectrum inhibitors of the metallo-
RT beta-lactamases.";
RL J. Med. Chem. 53:4862-4876(2010).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (2.03 ANGSTROMS) OF 28-254.
RA Delbruck H., Bebrone C., Hoffmann K.M.V., Galleni M.;
RT "Crystal structure of mutant C221D of carbapenemase CphA from Aeromonas
RT hydrophila.";
RL Submitted (AUG-2011) to the PDB data bank.
CC -!- FUNCTION: Confers resistance to the different beta-lactams antibiotics
CC (penicillin, cephalosporin and carbapenem) via the hydrolysis of the
CC beta-lactam ring (PubMed:15588826, PubMed:17307979, PubMed:1856163,
CC PubMed:19651913, PubMed:2327760). It is able to hydrolyze penicillin
CC and imipenem, but is much less active against cephalothin, cefotaxime,
CC meropenem and ceftazidime (PubMed:1856163, PubMed:2327760).
CC {ECO:0000269|PubMed:15588826, ECO:0000269|PubMed:17307979,
CC ECO:0000269|PubMed:1856163, ECO:0000269|PubMed:19651913,
CC ECO:0000269|PubMed:2327760}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a beta-lactam + H2O = a substituted beta-amino acid;
CC Xref=Rhea:RHEA:20401, ChEBI:CHEBI:15377, ChEBI:CHEBI:35627,
CC ChEBI:CHEBI:140347; EC=3.5.2.6;
CC Evidence={ECO:0000269|PubMed:15588826, ECO:0000269|PubMed:17307979,
CC ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:2327760};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:15588826, ECO:0000269|PubMed:17307979,
CC ECO:0000269|PubMed:18563261, ECO:0000269|PubMed:19651913,
CC ECO:0000269|PubMed:20527888};
CC Note=Binds 1 Zn(2+) ions per subunit. {ECO:0000269|PubMed:15588826,
CC ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:18563261,
CC ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:20527888};
CC -!- ACTIVITY REGULATION: Competitively inhibited by mercaptophosphonate and
CC pyridine carboxylate derivatives (PubMed:17307979, PubMed:20527888).
CC Also inhibited by the binding of a second zinc ion and by chelating
CC agents such as EDTA (PubMed:1856163, PubMed:2327760, PubMed:19651913).
CC {ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:1856163,
CC ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:20527888,
CC ECO:0000269|PubMed:2327760}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.8 uM for cefuroxime {ECO:0000269|PubMed:2327760};
CC KM=9 uM for ceftazidime {ECO:0000269|PubMed:2327760};
CC KM=15 uM for cefotaxime {ECO:0000269|PubMed:2327760};
CC KM=19 uM for cloxacillin {ECO:0000269|PubMed:2327760};
CC KM=28 uM for cephalothin {ECO:0000269|PubMed:2327760};
CC KM=30 uM for imipenem {ECO:0000269|PubMed:2327760};
CC KM=30.6 uM for nitrocefin {ECO:0000269|PubMed:2327760};
CC KM=53.3 uM for aztreonam {ECO:0000269|PubMed:2327760};
CC KM=63 uM for imipenem (at pH 9) {ECO:0000269|PubMed:17307979};
CC KM=83 uM for imipenem (at pH 8) {ECO:0000269|PubMed:17307979};
CC KM=120 uM for imipenem (at pH 7) {ECO:0000269|PubMed:17307979};
CC KM=124 uM for penicillin G {ECO:0000269|PubMed:2327760};
CC KM=166 uM for biapenem {ECO:0000269|PubMed:15588826};
CC KM=340 uM for imipenem {ECO:0000269|PubMed:15588826,
CC ECO:0000269|PubMed:19651913};
CC KM=1300 uM for nitrocefin {ECO:0000269|PubMed:15588826,
CC ECO:0000269|PubMed:19651913};
CC Vmax=170 umol/min/mg enzyme with penicillin G as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=100 umol/min/mg enzyme with nitrocefin as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=68.5 umol/min/mg enzyme with imipenem as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=12.8 umol/min/mg enzyme with cefuroxime as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=3.2 umol/min/mg enzyme with aztreonam as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=1.8 umol/min/mg enzyme with cephalothin as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=1.1 umol/min/mg enzyme with cloxacillin as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=0.5 umol/min/mg enzyme with cefotaxime as substrate
CC {ECO:0000269|PubMed:2327760};
CC Vmax=0.3 umol/min/mg enzyme with ceftazidime as substrate
CC {ECO:0000269|PubMed:2327760};
CC Note=kcat is 1200 sec(-1) for lactamase activity with imipenem as
CC substrate (PubMed:15588826, PubMed:19651913). kcat is 300 sec(-1) for
CC lactamase activity with biapenem as substrate (PubMed:15588826). kcat
CC is 210 sec(-1) for lactamase activity with imipenem as substrate (at
CC pH 7) (PubMed:17307979). kcat is 140 sec(-1) for lactamase activity
CC with imipenem as substrate (at pH 8) (PubMed:17307979). kcat is 49
CC sec(-1) for lactamase activity with imipenem as substrate (at pH 9)
CC (PubMed:17307979). kcat is 0.008 sec(-1) for lactamase activity with
CC nitrocefin as substrate (PubMed:19651913).
CC {ECO:0000269|PubMed:15588826, ECO:0000269|PubMed:17307979,
CC ECO:0000269|PubMed:19651913};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:15588826,
CC ECO:0000269|PubMed:1856163, ECO:0000269|PubMed:2327760}.
CC -!- SUBCELLULAR LOCATION: Periplasm {ECO:0000305|PubMed:1856163}.
CC -!- INDUCTION: By either cefoxitin or imipenem.
CC {ECO:0000269|PubMed:2327760}.
CC -!- SIMILARITY: Belongs to the metallo-beta-lactamase superfamily. Class-B
CC beta-lactamase family. {ECO:0000305}.
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DR EMBL; X57102; CAA40386.1; -; Genomic_DNA.
DR PIR; S17287; S17287.
DR RefSeq; WP_063844282.1; NG_047667.1.
DR PDB; 1X8G; X-ray; 1.70 A; A=28-254.
DR PDB; 1X8H; X-ray; 1.60 A; A=28-254.
DR PDB; 1X8I; X-ray; 1.90 A; A=28-254.
DR PDB; 2GKL; X-ray; 1.86 A; A=28-254.
DR PDB; 2QDS; X-ray; 1.66 A; A=28-254.
DR PDB; 3F9O; X-ray; 2.03 A; A=28-254.
DR PDB; 3FAI; X-ray; 1.70 A; A=28-254.
DR PDB; 3IOF; X-ray; 1.44 A; A=28-254.
DR PDB; 3IOG; X-ray; 1.41 A; A=28-254.
DR PDB; 3SW3; X-ray; 2.35 A; A=28-254.
DR PDB; 3T9M; X-ray; 2.03 A; A=28-254.
DR PDBsum; 1X8G; -.
DR PDBsum; 1X8H; -.
DR PDBsum; 1X8I; -.
DR PDBsum; 2GKL; -.
DR PDBsum; 2QDS; -.
DR PDBsum; 3F9O; -.
DR PDBsum; 3FAI; -.
DR PDBsum; 3IOF; -.
DR PDBsum; 3IOG; -.
DR PDBsum; 3SW3; -.
DR PDBsum; 3T9M; -.
DR AlphaFoldDB; P26918; -.
DR SMR; P26918; -.
DR BindingDB; P26918; -.
DR ChEMBL; CHEMBL1169593; -.
DR DrugBank; DB02032; Epicaptopril.
DR BRENDA; 3.5.2.6; 164.
DR SABIO-RK; P26918; -.
DR EvolutionaryTrace; P26918; -.
DR GO; GO:0042597; C:periplasmic space; IEA:UniProtKB-SubCell.
DR GO; GO:0008800; F:beta-lactamase activity; IDA:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0017001; P:antibiotic catabolic process; IDA:UniProtKB.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR Gene3D; 3.60.15.10; -; 1.
DR InterPro; IPR001018; Beta-lactamase_class-B_CS.
DR InterPro; IPR001279; Metallo-B-lactamas.
DR InterPro; IPR036866; RibonucZ/Hydroxyglut_hydro.
DR Pfam; PF00753; Lactamase_B; 1.
DR SMART; SM00849; Lactamase_B; 1.
DR SUPFAM; SSF56281; SSF56281; 1.
DR PROSITE; PS00743; BETA_LACTAMASE_B_1; 1.
DR PROSITE; PS00744; BETA_LACTAMASE_B_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; Hydrolase; Metal-binding; Periplasm;
KW Signal; Zinc.
FT SIGNAL 1..27
FT /evidence="ECO:0000255"
FT CHAIN 28..254
FT /note="Metallo-beta-lactamase type 2"
FT /id="PRO_0000016941"
FT BINDING 99
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:18563261,
FT ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:20527888"
FT BINDING 135
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15588826"
FT BINDING 174
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:18563261, ECO:0000269|PubMed:20527888"
FT BINDING 193
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:18563261,
FT ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:20527888"
FT BINDING 196
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:20527888"
FT BINDING 201
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:18563261"
FT BINDING 231
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:18563261,
FT ECO:0000269|PubMed:19651913, ECO:0000269|PubMed:20527888"
FT MUTAGEN 93
FT /note="N->H: Behaves as B1 and B3 enzymes, which are more
FT inhibited by 2-picolinic acid than by 2,4-PDCA; when
FT associated with G-192."
FT /evidence="ECO:0000269|PubMed:17307979"
FT MUTAGEN 192
FT /note="N->G: Slight decrease of the catalytic efficiency
FT and slight increase of the affinity for both biapenem and
FT imipenem. Behaves as B1 and B3 enzymes, which are more
FT inhibited by 2-picolinic acid than by 2,4-PDCA; when
FT associated with H-93."
FT /evidence="ECO:0000269|PubMed:15588826,
FT ECO:0000269|PubMed:17307979, ECO:0000269|PubMed:19651913"
FT STRAND 30..36
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 39..44
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 46..48
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 50..56
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 61..65
FT /evidence="ECO:0007829|PDB:3IOG"
FT HELIX 70..81
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 88..92
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 94..97
FT /evidence="ECO:0007829|PDB:3IOG"
FT HELIX 98..101
FT /evidence="ECO:0007829|PDB:3IOG"
FT HELIX 104..109
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 113..117
FT /evidence="ECO:0007829|PDB:3IOG"
FT HELIX 118..138
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 151..156
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 158..160
FT /evidence="ECO:0007829|PDB:3IOG"
FT TURN 161..164
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 165..168
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 173..177
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 180..183
FT /evidence="ECO:0007829|PDB:3IOG"
FT TURN 184..187
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 188..190
FT /evidence="ECO:0007829|PDB:3IOG"
FT HELIX 192..194
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 198..200
FT /evidence="ECO:0007829|PDB:3SW3"
FT HELIX 209..219
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 225..228
FT /evidence="ECO:0007829|PDB:3IOG"
FT STRAND 230..232
FT /evidence="ECO:0007829|PDB:3IOG"
FT HELIX 239..250
FT /evidence="ECO:0007829|PDB:3IOG"
SQ SEQUENCE 254 AA; 28016 MW; 9D54760E17CF4B50 CRC64;
MMKGWMKCGL AGAVVLMASF WGGSVRAAGM SLTQVSGPVY VVEDNYYVQE NSMVYFGAKG
VTVVGATWTP DTARELHKLI KRVSRKPVLE VINTNYHTDR AGGNAYWKSI GAKVVSTRQT
RDLMKSDWAE IVAFTRKGLP EYPDLPLVLP NVVHDGDFTL QEGKVRAFYA GPAHTPDGIF
VYFPDEQVLY GNCILKEKLG NLSFADVKAY PQTLERLKAM KLPIKTVIGG HDSPLHGPEL
IDHYEALIKA APQS
