PEPT_LACHE
ID PEPT_LACHE Reviewed; 413 AA.
AC Q9L4G1;
DT 02-MAY-2002, integrated into UniProtKB/Swiss-Prot.
DT 03-APR-2007, sequence version 2.
DT 03-AUG-2022, entry version 89.
DE RecName: Full=Peptidase T;
DE EC=3.4.11.4;
DE AltName: Full=Aminotripeptidase;
DE Short=Tripeptidase;
DE AltName: Full=Tripeptide aminopeptidase;
GN Name=pepT;
OS Lactobacillus helveticus (Lactobacillus suntoryeus).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae;
OC Lactobacillus.
OX NCBI_TaxID=1587;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-17, FUNCTION,
RP CHARACTERIZATION, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION, SUBUNIT, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=53/7;
RX PubMed=10653753; DOI=10.1128/aem.66.2.794-800.2000;
RA Savijoki K., Palva A.;
RT "Purification and molecular characterization of a tripeptidase from
RT Lactobacillus helveticus.";
RL Appl. Environ. Microbiol. 66:794-800(2000).
CC -!- FUNCTION: Cleaves the N-terminal amino acid of tripeptides. Shows broad
CC substrate specificity, exhibiting maximum activity against hydrophobic
CC tripeptides, with the highest activity for Met-Gly-Gly. Therefore this
CC enzyme may play an important role in flavor formation during cheese
CC ripening. Is also able to slowly hydrolyze some hydrophobic dipeptides,
CC but displays no activity against tetrapeptides and the tripeptide Phe-
CC Gly-Gly. {ECO:0000269|PubMed:10653753}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Release of the N-terminal residue from a tripeptide.;
CC EC=3.4.11.4;
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105; Evidence={ECO:0000305};
CC Note=Binds 2 Zn(2+) ions per subunit. {ECO:0000305};
CC -!- ACTIVITY REGULATION: Totally inhibited by EDTA, EGTA, and 1,10-
CC phenanthroline. Strongly inhibited by divalent cations such as Cu(2+),
CC Cd(2+), Co(2+) and Mn(2+). Partially inhibited by the reducing agents
CC 2-mercaptoethanol and dithiothreitol. {ECO:0000269|PubMed:10653753}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.6 mM for tripeptide Met-Gly-Gly {ECO:0000269|PubMed:10653753};
CC KM=0.6 mM for tripeptide Leu-Gly-Gly {ECO:0000269|PubMed:10653753};
CC Vmax=80.2 umol/min/ug enzyme with Met-Gly-Gly as substrate
CC {ECO:0000269|PubMed:10653753};
CC Vmax=6.8 umol/min/ug enzyme with Leu-Gly-Gly as substrate
CC {ECO:0000269|PubMed:10653753};
CC pH dependence:
CC Optimum pH is 7.5. Highly active from pH 6 to 8.
CC {ECO:0000269|PubMed:10653753};
CC Temperature dependence:
CC Optimum temperature is 25-37 degrees Celsius. Thermolabile.
CC Approximately 45% of the activity remains after incubation for 15 min
CC at 45 degrees Celsius. {ECO:0000269|PubMed:10653753};
CC -!- SUBUNIT: Homotrimer. {ECO:0000305|PubMed:10653753}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the peptidase M20B family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CAB72938.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; AJ243321; CAB72938.1; ALT_FRAME; Genomic_DNA.
DR AlphaFoldDB; Q9L4G1; -.
DR SMR; Q9L4G1; -.
DR STRING; 326425.lhe_1170; -.
DR eggNOG; COG2195; Bacteria.
DR BRENDA; 3.4.11.4; 2870.
DR SABIO-RK; Q9L4G1; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0008237; F:metallopeptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0045148; F:tripeptide aminopeptidase activity; IEA:UniProtKB-EC.
DR GO; GO:0008270; F:zinc ion binding; IEA:InterPro.
DR GO; GO:0006518; P:peptide metabolic process; IEA:InterPro.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
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; PS00759; ARGE_DAPE_CPG2_2; 1.
PE 1: Evidence at protein level;
KW Aminopeptidase; Cytoplasm; Direct protein sequencing; Hydrolase;
KW Metal-binding; Metalloprotease; Protease; Zinc.
FT CHAIN 1..413
FT /note="Peptidase T"
FT /id="PRO_0000185300"
FT ACT_SITE 84
FT /evidence="ECO:0000250"
FT ACT_SITE 178
FT /note="Proton acceptor"
FT /evidence="ECO:0000250"
FT BINDING 82
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 144
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 144
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 179
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 201
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 383
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
SQ SEQUENCE 413 AA; 46681 MW; 704E49E8480D2835 CRC64;
MEYPNLLPRF LKYVKVNSRS DENSDRFPST EREENFQKNV IMKDLEELGL SDIHYNQKAG
SVIAEIPSNV DYDVPVMGFL AHSDTADFNS ENVKPQIHKN YDGESKIQLG DSEFYLDPEV
YPNLRKYKGQ TIITASGDTL LGADDKCGIS ELMTFAEYLM NHPEVKHGKI RLAFTPDEEI
GTGAEQFDVK DFGADFAFTV DGEAPGKLGD CTFSAAQFTL DIQGVNVHPA VAKGQMINAV
QVGIDFHNQL PEHDRPEHTD GREGFFHLLS FDGTVDHAHL AYIIRDFERD GLEERKNLVK
SIVKKMNDEF GTERIKLQMN DQYYNMADEL KKHMDIVDLA RDAYKAEGLE VNEDPVRGGT
DGSQLTYMGL PCPNIFAGEE NMHGRYEYTV LESMYKTVDV MIKMAELNAE RAK
