CBP1_THEAQ
ID CBP1_THEAQ Reviewed; 511 AA.
AC P42663;
DT 01-NOV-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1996, sequence version 2.
DT 03-AUG-2022, entry version 89.
DE RecName: Full=Thermostable carboxypeptidase 1;
DE EC=3.4.17.19;
DE AltName: Full=Carboxypeptidase Taq;
OS Thermus aquaticus.
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=271;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], CATALYTIC ACTIVITY, COFACTOR, FUNCTION,
RP AND ACTIVITY REGULATION.
RC STRAIN=ATCC 25104 / DSM 625 / JCM 10724 / NBRC 103206 / NCIMB 11243 / YT-1;
RX PubMed=7765282; DOI=10.1271/bbb.58.1490;
RA Lee S.-H., Taguchi H., Yoshimura E., Minagawa E., Kaminogawa S., Ohta T.,
RA Matsuzawa H.;
RT "Carboxypeptidase Taq, a thermostable zinc enzyme, from Thermus aquaticus
RT YT-1: molecular cloning, sequencing, and expression of the encoding gene in
RT Escherichia coli.";
RL Biosci. Biotechnol. Biochem. 58:1490-1495(1994).
RN [2]
RP PROTEIN SEQUENCE OF 1-28, CATALYTIC ACTIVITY, COFACTOR, FUNCTION, ACTIVITY
RP REGULATION, SUBUNIT, AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=ATCC 25104 / DSM 625 / JCM 10724 / NBRC 103206 / NCIMB 11243 / YT-1;
RX PubMed=1369078; DOI=10.1271/bbb.56.1839;
RA Lee S.-H., Minagawa E., Taguchi H., Matsuzawa H., Ohta T., Kaminogawa S.,
RA Yamauchi K.;
RT "Purification and characterization of a thermostable carboxypeptidase
RT (carboxypeptidase Taq) from Thermus aquaticus YT-1.";
RL Biosci. Biotechnol. Biochem. 56:1839-1844(1992).
RN [3]
RP METAL BINDING SITES, MUTAGENESIS OF HIS-276; GLU-277; HIS-280; GLU-298 AND
RP GLU-332, COFACTOR, ACTIVE SITE, AND CATALYTIC ACTIVITY.
RX PubMed=8862545; DOI=10.1093/protein/9.6.467;
RA Lee S.H., Taguchi H., Yoshimura E., Minagawa E., Kaminogawa S., Ohta T.,
RA Matsuzawa H.;
RT "The active site of carboxypeptidase Taq possesses the active-site motif
RT His-Glu-X-X-His of zinc-dependent endopeptidases and aminopeptidases.";
RL Protein Eng. 9:467-469(1996).
CC -!- FUNCTION: Broad specificity carboxypetidase that releases amino acids
CC sequentially from the C-terminus, including neutral, aromatic, polar
CC and basic residues, but not Pro. {ECO:0000269|PubMed:1369078,
CC ECO:0000269|PubMed:7765282}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Release of a C-terminal amino acid with broad specificity,
CC except for -Pro.; EC=3.4.17.19; Evidence={ECO:0000269|PubMed:1369078,
CC ECO:0000269|PubMed:7765282, ECO:0000269|PubMed:8862545};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:1369078, ECO:0000269|PubMed:7765282,
CC ECO:0000269|PubMed:8862545};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:1369078, ECO:0000269|PubMed:7765282,
CC ECO:0000269|PubMed:8862545};
CC Note=Binds 1 zinc ion per subunit. Can also utilize cobalt ions, but
CC contains bound zinc when purified after heterologous expression in
CC E.coli. {ECO:0000269|PubMed:1369078, ECO:0000269|PubMed:7765282,
CC ECO:0000269|PubMed:8862545};
CC -!- ACTIVITY REGULATION: Inhibited by metal-chelating reagents. Strongly
CC inhibited by 1 mM Zn(2+), slightly inhibited by 0.1 mM Zn(2+), not
CC inhibited by 0.01 mM Zn(2+). Not inhibited by cobalt ions.
CC {ECO:0000269|PubMed:1369078, ECO:0000269|PubMed:7765282}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:1369078};
CC Temperature dependence:
CC Optimum temperature is 80 degrees Celsius.
CC {ECO:0000269|PubMed:1369078};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:1369078}.
CC -!- SIMILARITY: Belongs to the peptidase M32 family. {ECO:0000305}.
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DR EMBL; D17669; BAA04551.1; -; Genomic_DNA.
DR PIR; JC2294; JC2294.
DR RefSeq; WP_053767417.1; NZ_LHCI01000106.1.
DR AlphaFoldDB; P42663; -.
DR SMR; P42663; -.
DR MEROPS; M32.001; -.
DR KEGG; ag:BAA04551; -.
DR BRENDA; 3.4.17.19; 6334.
DR GO; GO:0004181; F:metallocarboxypeptidase activity; IDA:UniProtKB.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IDA:UniProtKB.
DR InterPro; IPR001333; Peptidase_M32_Taq.
DR PANTHER; PTHR34217; PTHR34217; 1.
DR Pfam; PF02074; Peptidase_M32; 1.
DR PIRSF; PIRSF006615; Zn_crbxpep_Taq; 1.
DR PRINTS; PR00998; CRBOXYPTASET.
DR PROSITE; PS00142; ZINC_PROTEASE; 1.
PE 1: Evidence at protein level;
KW Carboxypeptidase; Cobalt; Direct protein sequencing; Hydrolase;
KW Metal-binding; Metalloprotease; Protease; Zinc; Zymogen.
FT CHAIN 1..511
FT /note="Thermostable carboxypeptidase 1"
FT /id="PRO_0000078235"
FT ACT_SITE 277
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000305|PubMed:8862545"
FT BINDING 276
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:8862545"
FT BINDING 280
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:8862545"
FT BINDING 306
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10095,
FT ECO:0000269|PubMed:8862545"
FT MUTAGEN 276
FT /note="H->E: Reduces enzyme activity. No effect on zinc
FT binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 276
FT /note="H->R,Y: Reduces enzyme activity. Impairs zinc
FT binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 276
FT /note="H->R: Reduces enzyme activity. Impairs zinc
FT binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 277
FT /note="E->D,Q: Reduces enzyme activity. No effect on zinc
FT binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 280
FT /note="H->E: Reduces enzyme activity. Impairs zinc
FT binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 280
FT /note="H->R,Y: Reduces enzyme activity. Impairs zinc
FT binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 298
FT /note="E->Q: No effect on enzyme activity. Slightly reduces
FT zinc binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT MUTAGEN 332
FT /note="E->Q: No effect on enzyme activity. Slightly reduces
FT zinc binding."
FT /evidence="ECO:0000269|PubMed:8862545"
FT CONFLICT 9
FT /note="N -> D (in Ref. 2; AA sequence)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 511 AA; 57954 MW; 659DF4EBCDEE3ABD CRC64;
MTPEAAYQNL LEFQRETAYL GSLGALAAWD QRTMIPRKGH GHRARQMAAL ARLLHERATD
PRIGEWLEKV EGSSLVEDPL SDAAVNVRAW RRAYERARAI PERLAVELAQ ARSEGETAWE
ALRPRDDWQG FLPYLKRLFA LAKEEAEILM AVGPDPLDPP YGELYDALLD GYEPGARARD
LEPLFRELSS GLKGLLDRIL GSGRRPDVGV LHRHYPKEAQ RAFALELLQA CGYDLEAGRL
DPTAHPFEIA IGPGDVRITT RYYEDFFNAG IFGTLHEMGH ALYEQGLPEA HWGTPRGEAA
SLGVHESQSR TWENLVGRSL GFWERFFPRA KEVFSSLADV RLEDFHFAVN AVEPSLIRVE
ADEVTYNLHI LVRLELELAL FRGELFLEDL PEAWREKYRA YLGVAPRDYK DGVMQDVHWS
GGMFGYFPTY TLGNLYAAQF FAKAQEELGP LEPLFARGEF TPFLDWTRRK IHAEGSRFRP
RALVERVTGS PPGAQAFLRY LEAKYGALYG F
