PEPT_LACLL
ID PEPT_LACLL Reviewed; 413 AA.
AC Q76HM5;
DT 07-JAN-2015, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 109.
DE RecName: Full=Peptidase T {ECO:0000255|HAMAP-Rule:MF_00550};
DE EC=3.4.11.4 {ECO:0000255|HAMAP-Rule:MF_00550, ECO:0000269|PubMed:15752689, ECO:0000269|PubMed:16233215};
DE AltName: Full=Aminotripeptidase {ECO:0000255|HAMAP-Rule:MF_00550, ECO:0000303|PubMed:16233215};
DE Short=Tripeptidase {ECO:0000255|HAMAP-Rule:MF_00550, ECO:0000303|PubMed:15752689, ECO:0000303|PubMed:16233215};
DE AltName: Full=L9PepTR {ECO:0000303|PubMed:15752689};
DE AltName: Full=Tripeptide aminopeptidase {ECO:0000255|HAMAP-Rule:MF_00550};
GN Name=pepT {ECO:0000255|HAMAP-Rule:MF_00550, ECO:0000303|PubMed:15752689};
OS Lactococcus lactis subsp. lactis (Streptococcus lactis).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Lactococcus.
OX NCBI_TaxID=1360;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 13675 / LMG 9441 / NCDO 1007, NIAI N-7, NIRD DRC-1, and
RC NIRD DRC-2;
RX PubMed=15368846; DOI=10.1078/0723202041438400;
RA Mori S., Mori K., Suzuki I., Kasumi T.;
RT "Phylogenetic analysis of Lactococcus lactis subspecies based on decoding
RT the sequence of the pepT tripeptidase gene, the pepV dipeptidase gene and
RT 16S rRNA.";
RL Syst. Appl. Microbiol. 27:414-422(2004).
RN [2]
RP PROTEIN SEQUENCE OF N-TERMINUS, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, AND SUBUNIT.
RC STRAIN=ATCC 13675 / LMG 9441 / NCDO 1007;
RX PubMed=15752689; DOI=10.1016/j.bbapap.2004.12.001;
RA Mori S., Nirasawa S., Komba S., Kasumi T.;
RT "Characterization and kinetic analysis of enzyme-substrate recognition by
RT three recombinant lactococcal tripeptidases.";
RL Biochim. Biophys. Acta 1748:26-34(2005).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBSTRATE SPECIFICITY, ACTIVITY REGULATION, AND SUBUNIT.
RC STRAIN=ATCC 13675 / LMG 9441 / NCDO 1007;
RX PubMed=16233215; DOI=10.1016/s1389-1723(02)80068-8;
RA Mori S., Sumino S., Kasumi T.;
RT "Substrate specificity of a tripeptidase as a metalloenzyme purified from
RT Lactococcus lactis subsp. lactis biovar. diacetylactis ATCC 13675.";
RL J. Biosci. Bioeng. 93:360-366(2002).
CC -!- FUNCTION: Cleaves the N-terminal amino acid of tripeptides.
CC {ECO:0000255|HAMAP-Rule:MF_00550, ECO:0000269|PubMed:15752689,
CC ECO:0000269|PubMed:16233215}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Release of the N-terminal residue from a tripeptide.;
CC EC=3.4.11.4; Evidence={ECO:0000255|HAMAP-Rule:MF_00550,
CC ECO:0000269|PubMed:15752689, ECO:0000269|PubMed:16233215};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00550,
CC ECO:0000269|PubMed:16233215};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:16233215};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:16233215};
CC Note=Binds 2 Zn(2+) ions per subunit. To a lesser extent, can also use
CC Co(2+) or Mn(2+) instead of Zn(2+). {ECO:0000255|HAMAP-Rule:MF_00550,
CC ECO:0000269|PubMed:16233215};
CC -!- ACTIVITY REGULATION: Activity is strongly inhibited in vitro by
CC chelating agents such as EDTA and 1,10-phenanthroline as do the
CC reducing agent dithiothreitol and the oxidizing agent N-
CC bromosuccinimid. Phenylmethylsulfonyl fluoride, a serine protease
CC inhibitor has no influence on the enzyme activity. On the other hand,
CC bestatin, an aminopeptidase inhibitor strongly inhibits the enzyme
CC activity and leupeptin, a modified tripeptide, slightly inhibits it.
CC {ECO:0000269|PubMed:16233215}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.147 mM for AAA tripeptide {ECO:0000269|PubMed:15752689};
CC KM=0.390 mM for GAA tripeptide {ECO:0000269|PubMed:15752689};
CC KM=0.598 mM for AGA tripeptide {ECO:0000269|PubMed:15752689};
CC KM=1.29 mM for AAG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=0.770 mM for GGA tripeptide {ECO:0000269|PubMed:15752689};
CC KM=1.57 mM for AGG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=4.52 mM for GAG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=12.0 mM for GGG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=1.77 mM for YGG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=2.44 mM for LGG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=3.54 mM for FGG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=6.49 mM for SGG tripeptide {ECO:0000269|PubMed:15752689};
CC KM=10.6 mM for AibGG tripeptide {ECO:0000269|PubMed:15752689};
CC Note=kcat is 163 sec(-1) with AAA tripeptide as substrate. kcat is
CC 616 sec(-1) with GAA tripeptide as substrate. kcat is 713 sec(-1)
CC with AGA tripeptide as substrate. kcat is 730 sec(-1) with AAG
CC tripeptide as substrate. kcat is 507 sec(-1) with GGA tripeptide as
CC substrate. kcat is 573 sec(-1) with AGG tripeptide as substrate. kcat
CC is 1093 sec(-1) with GAG tripeptide as substrate. kcat is 502 sec(-1)
CC with GGG tripeptide as substrate. kcat is 292 sec(-1) with YGG
CC tripeptide as substrate. kcat is 197 sec(-1) with LGG tripeptide as
CC substrate. kcat is 262 sec(-1) with FGG tripeptide as substrate. kcat
CC is 270 sec(-1) with SGG tripeptide as substrate. kcat is 31.2 sec(-1)
CC with AibGG tripeptide as substrate. kcat is 453 sec(-1) with GGF
CC tripeptide as substrate. kcat is 806 sec(-1) with GAY tripeptide as
CC substrate. {ECO:0000269|PubMed:15752689};
CC pH dependence:
CC Optimum pH is around 8. The enzyme is stable in a pH range of 9-10.
CC {ECO:0000269|PubMed:15752689, ECO:0000269|PubMed:16233215};
CC Temperature dependence:
CC Optimum temperature is around 44 degrees Celsius. The enzyme is
CC stable up to 50 degrees Celsius. {ECO:0000269|PubMed:15752689,
CC ECO:0000269|PubMed:16233215};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:15752689,
CC ECO:0000269|PubMed:16233215}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_00550}.
CC -!- SIMILARITY: Belongs to the peptidase M20B family. {ECO:0000255|HAMAP-
CC Rule:MF_00550}.
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DR EMBL; AB100769; BAC66669.1; -; Genomic_DNA.
DR EMBL; AB100770; BAC66670.1; -; Genomic_DNA.
DR EMBL; AB100771; BAC66671.1; -; Genomic_DNA.
DR EMBL; AB100775; BAC66675.1; -; Genomic_DNA.
DR RefSeq; WP_003130321.1; NZ_WJUU01000017.1.
DR AlphaFoldDB; Q76HM5; -.
DR SMR; Q76HM5; -.
DR GeneID; 60355196; -.
DR OMA; GHNFHGK; -.
DR SABIO-RK; Q76HM5; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0050897; F:cobalt ion binding; IDA:UniProtKB.
DR GO; GO:0030145; F:manganese ion binding; IDA:UniProtKB.
DR GO; GO:0008237; F:metallopeptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:0045148; F:tripeptide aminopeptidase activity; IDA:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0043171; P:peptide catabolic process; IEA:UniProtKB-UniRule.
DR GO; GO:0006518; P:peptide metabolic process; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IDA:UniProtKB.
DR HAMAP; MF_00550; Aminopeptidase_M20; 1.
DR InterPro; IPR001261; ArgE/DapE_CS.
DR InterPro; IPR036264; Bact_exopeptidase_dim_dom.
DR InterPro; IPR002933; Peptidase_M20.
DR InterPro; IPR011650; Peptidase_M20_dimer.
DR InterPro; IPR010161; Peptidase_M20B.
DR PANTHER; PTHR42994:SF1; PTHR42994:SF1; 1.
DR Pfam; PF07687; M20_dimer; 1.
DR Pfam; PF01546; Peptidase_M20; 1.
DR PIRSF; PIRSF037215; Peptidase_M20B; 1.
DR SUPFAM; SSF55031; SSF55031; 1.
DR TIGRFAMs; TIGR01882; peptidase-T; 1.
DR PROSITE; PS00758; ARGE_DAPE_CPG2_1; 1.
DR PROSITE; PS00759; ARGE_DAPE_CPG2_2; 1.
PE 1: Evidence at protein level;
KW Aminopeptidase; Cobalt; Cytoplasm; Direct protein sequencing; Hydrolase;
KW Manganese; Metal-binding; Metalloprotease; Protease; Zinc.
FT CHAIN 1..413
FT /note="Peptidase T"
FT /id="PRO_0000431585"
FT ACT_SITE 83
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT ACT_SITE 178
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT BINDING 81
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT BINDING 143
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT BINDING 143
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT BINDING 179
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT BINDING 201
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
FT BINDING 383
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00550"
SQ SEQUENCE 413 AA; 45972 MW; 52E26C8E7F518FEC CRC64;
MKYEKLLPRF LEYVKVNTRS DENSTTTPST QALVEFAHKM GEDMKALGLK DVHYLESNGY
VIGTIPANTD KKVRKIGLLA HLDTADFNAE GVNPQILENY DGESVIKLGD TEFTLDPKDF
PNLKNYKGQT LVHTDGTTLL GSDDKSGVAE IMTLADYLLN INPDFEHGEI RVGFGPDEEI
GVGADKFDVA DFDVDFAYTV DGGPLGELQY ETFSAAGAVI EFQGKNVHPG TAKNTMVNAL
QLAIDYHNAL PEFDRPEKTE GREGFFHLLK LDGTPEEARA QYIIRDHEEG KFNERKALMQ
EIADKMNAEL GQNRVKPVIK DQYYNMAQII EKDMSIIDIA KKAMENLDIV PIIEPIRGGT
DGSKISFMGL PTPNLFAGGE NMHGRFEFVS VQTMEKAVDT LLEIIRLNNE VVK
