COLT_CLOTE
ID COLT_CLOTE Reviewed; 991 AA.
AC Q899Y1;
DT 28-MAR-2018, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2003, sequence version 1.
DT 03-AUG-2022, entry version 97.
DE RecName: Full=Collagenase ColT {ECO:0000303|PubMed:12552129};
DE EC=3.4.24.3 {ECO:0000269|PubMed:24125730};
DE AltName: Full=Microbial collagenase;
DE Flags: Precursor;
GN Name=colT {ECO:0000303|PubMed:12552129}; OrderedLocusNames=CTC_p33;
OS Clostridium tetani (strain Massachusetts / E88).
OG Plasmid pE88.
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae;
OC Clostridium.
OX NCBI_TaxID=212717;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=Massachusetts / E88;
RX PubMed=12552129; DOI=10.1073/pnas.0335853100;
RA Brueggemann H., Baeumer S., Fricke W.F., Wiezer A., Liesegang H.,
RA Decker I., Herzberg C., Martinez-Arias R., Merkl R., Henne A.,
RA Gottschalk G.;
RT "The genome sequence of Clostridium tetani, the causative agent of tetanus
RT disease.";
RL Proc. Natl. Acad. Sci. U.S.A. 100:1316-1321(2003).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND DOMAIN.
RX PubMed=18937627; DOI=10.1515/bc.2009.004;
RA Eckhard U., Schoenauer E., Ducka P., Briza P., Nuess D., Brandstetter H.;
RT "Biochemical characterization of the catalytic domains of three different
RT Clostridial collagenases.";
RL Biol. Chem. 390:11-18(2009).
RN [3]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=24125730; DOI=10.1016/j.jprot.2013.10.004;
RA Eckhard U., Huesgen P.F., Brandstetter H., Overall C.M.;
RT "Proteomic protease specificity profiling of clostridial collagenases
RT reveals their intrinsic nature as dedicated degraders of collagen.";
RL J. Proteomics 100:102-114(2014).
RN [4]
RP FUNCTION, ACTIVITY REGULATION, AND BIOTECHNOLOGY FOR ANTI-INFECTIVE AGENTS.
RX PubMed=28820255; DOI=10.1021/jacs.7b06935;
RA Schoenauer E., Kany A.M., Haupenthal J., Huesecken K., Hoppe I.J., Voos K.,
RA Yahiaoui S., Elsaesser B., Ducho C., Brandstetter H., Hartmann R.W.;
RT "Discovery of a potent inhibitor class with high selectivity toward
RT clostridial collagenases.";
RL J. Am. Chem. Soc. 139:12696-12703(2017).
RN [5] {ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9}
RP X-RAY CRYSTALLOGRAPHY (1.69 ANGSTROMS) OF 340-730 IN PRESENCE OR ABSENCE OF
RP INHIBITOR AND IN COMPLEX WITH CALCIUM AND ZINC, FUNCTION, COFACTOR,
RP ACTIVITY REGULATION, AND DOMAIN.
RX PubMed=23703618; DOI=10.1074/jbc.m112.448548;
RA Eckhard U., Schonauer E., Brandstetter H.;
RT "Structural basis for activity regulation and substrate preference of
RT clostridial collagenases G, H, and T.";
RL J. Biol. Chem. 288:20184-20194(2013).
CC -!- FUNCTION: Clostridial collagenases are among the most efficient
CC degraders of eukaryotic collagen known; saprophytes use collagen as a
CC carbon source while pathogens additionally digest collagen to aid in
CC host colonization. Has both tripeptidylcarboxypeptidase on Gly-X-Y and
CC endopeptidase activities; the endopeptidase cuts within the triple
CC helix region of collagen while tripeptidylcarboxypeptidase successively
CC digests the exposed ends, thus clostridial collagenases can digest
CC large sections of collagen (By similarity). The activator domain
CC (residues 57-330) and catalytic subdomain (340-611) open and close
CC around substrate allowing digestion when the protein is closed
CC (PubMed:23703618). {ECO:0000250|UniProtKB:Q9X721,
CC ECO:0000269|PubMed:18937627, ECO:0000269|PubMed:23703618,
CC ECO:0000269|PubMed:24125730, ECO:0000269|PubMed:28820255}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Digestion of native collagen in the triple helical region at
CC Xaa-|-Gly bonds. With synthetic peptides, a preference is shown for
CC Gly at P3 and P1', Pro and Ala at P2 and P2', and hydroxyproline, Ala
CC or Arg at P3'.; EC=3.4.24.3; Evidence={ECO:0000269|PubMed:24125730,
CC ECO:0000305|PubMed:18937627, ECO:0000305|PubMed:23703618};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:23703618};
CC Note=Binds about 5 Ca(2+) per subunit (Probable). The metallopeptidase
CC domain binds 1 Ca(2+), while each CDB binds 2 (Probable).
CC {ECO:0000305};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:23703618};
CC Note=Binds 1 catalytic Zn(2+) per subunit.
CC {ECO:0000269|PubMed:23703618};
CC -!- ACTIVITY REGULATION: Partially inhibited by 1-10-phenanthroline;
CC inactivation is irreversible (PubMed:18937627, PubMed:23703618).
CC Partially inhibited by EDTA; inactivation is reversible
CC (PubMed:23703618). Inhibited by broad-spectrum zinc metalloprotease
CC inhibitor batimastat (PubMed:28820255). N-aryl mercaptoacetamide-based
CC inhibitors have been isolated that act on clostridial collagenases with
CC submicromolar affinity while having negligibile activity on human
CC collagenases (PubMed:28820255). {ECO:0000269|PubMed:18937627,
CC ECO:0000269|PubMed:23703618, ECO:0000269|PubMed:28820255}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.970 mM for furylacryloyl-Leu-Gly-Pro-Ala (FALGPA)
CC {ECO:0000269|PubMed:18937627};
CC Vmax=5.53 umol/min/mg enzyme {ECO:0000269|PubMed:18937627};
CC Note=kcat is 7.24/sec, using a catalytic fragment (residues 53-727)
CC on an artificial substrate. {ECO:0000269|PubMed:18937627};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250|UniProtKB:Q9X721}.
CC -!- DOMAIN: The mature protein has 3 domains; a metalloprotease domain (S1,
CC approximately residues 53 to 727) and 2 collagen-binding domains (CBD)
CC (755-872) and (882-991) (PubMed:18937627). The metalloprotease S1
CC domain is composed of 3 subdomains which together resemble a saddle; an
CC activator domain (residues 57-330), the catalytic peptidase subdomain
CC (340-611) and a helper subdomain (619-731) (PubMed:23703618).
CC {ECO:0000269|PubMed:23703618, ECO:0000305|PubMed:18937627}.
CC -!- BIOTECHNOLOGY: N-aryl mercaptoacetamide-based inhibitors with
CC submicromolar affinity for clostridial collagenases but negligibile
CC activity on human collagenases have been discovered that may lead to
CC promising anti-infective drugs against Clostridia (PubMed:28820255).
CC {ECO:0000269|PubMed:28820255}.
CC -!- SIMILARITY: Belongs to the peptidase M9B family. Collagenase subfamily.
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DR EMBL; AF528097; AAO37456.1; -; Genomic_DNA.
DR RefSeq; WP_011100838.1; NC_004565.1.
DR PDB; 4AR8; X-ray; 2.05 A; A/B=340-730.
DR PDB; 4AR9; X-ray; 1.69 A; A/B=340-730.
DR PDBsum; 4AR8; -.
DR PDBsum; 4AR9; -.
DR AlphaFoldDB; Q899Y1; -.
DR SMR; Q899Y1; -.
DR MEROPS; M09.005; -.
DR EnsemblBacteria; AAO37456; AAO37456; CTC_p33.
DR KEGG; ctc:CTC_p33; -.
DR HOGENOM; CLU_012279_0_0_9; -.
DR OMA; YEREGSY; -.
DR OrthoDB; 43759at2; -.
DR BRENDA; 3.4.24.3; 1526.
DR Proteomes; UP000001412; Plasmid pE88.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0005509; F:calcium ion binding; IDA:UniProtKB.
DR GO; GO:0004175; F:endopeptidase activity; IDA:UniProtKB.
DR GO; GO:0004222; F:metalloendopeptidase activity; IMP:UniProtKB.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:UniProtKB-EC.
DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR InterPro; IPR041379; ColG_subdomain.
DR InterPro; IPR007280; Peptidase_C_arc/bac.
DR InterPro; IPR013661; Peptidase_M9_N_dom.
DR InterPro; IPR002169; Peptidase_M9A/M9B.
DR Pfam; PF18496; ColG_sub; 1.
DR Pfam; PF01752; Peptidase_M9; 1.
DR Pfam; PF08453; Peptidase_M9_N; 1.
DR Pfam; PF04151; PPC; 2.
DR PRINTS; PR00931; MICOLLPTASE.
DR PROSITE; PS00142; ZINC_PROTEASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Hydrolase; Metal-binding; Metalloprotease; Plasmid;
KW Protease; Reference proteome; Repeat; Secreted; Signal; Virulence; Zinc;
KW Zymogen.
FT SIGNAL 1..28
FT /evidence="ECO:0000255"
FT PROPEP 29..52
FT /evidence="ECO:0000303|PubMed:18937627"
FT /id="PRO_0000443548"
FT CHAIN 53..991
FT /note="Collagenase ColT"
FT /id="PRO_5004302093"
FT REGION 53..727
FT /note="S1 metalloprotease domain, degrades FALGPA
FT (furylacryloyl-Leu-Gly-Pro-Ala)"
FT /evidence="ECO:0000305|PubMed:18937627,
FT ECO:0000305|PubMed:23703618"
FT REGION 57..330
FT /note="Activator domain"
FT /evidence="ECO:0000305|PubMed:23703618"
FT REGION 340..611
FT /note="Catalytic subdomain"
FT /evidence="ECO:0000305|PubMed:23703618"
FT REGION 619..731
FT /note="Helper subdomain"
FT /evidence="ECO:0000305|PubMed:23703618"
FT REGION 755..870
FT /note="Collagen-binding domain 1"
FT /evidence="ECO:0000305|PubMed:18937627,
FT ECO:0000305|PubMed:23703618"
FT REGION 878..991
FT /note="Collagen-binding domain 2"
FT /evidence="ECO:0000305|PubMed:18937627,
FT ECO:0000305|PubMed:23703618"
FT ACT_SITE 466
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10095"
FT BINDING 440
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23703618,
FT ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"
FT BINDING 465
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:23703618, ECO:0007744|PDB:4AR8,
FT ECO:0007744|PDB:4AR9"
FT BINDING 469
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:23703618, ECO:0007744|PDB:4AR8,
FT ECO:0007744|PDB:4AR9"
FT BINDING 473
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23703618,
FT ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"
FT BINDING 477
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23703618,
FT ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"
FT BINDING 479
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:23703618,
FT ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"
FT BINDING 499
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:23703618,
FT ECO:0007744|PDB:4AR8, ECO:0007744|PDB:4AR9"
FT BINDING 757
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 759
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 759
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 761
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 784
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 784
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 787
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 787
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 883
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="4"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 885
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="4"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 885
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="5"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 887
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="5"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 888
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="5"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 910
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="4"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 910
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="5"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 913
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="4"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT BINDING 913
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="5"
FT /evidence="ECO:0000250|UniProtKB:Q9X721"
FT HELIX 341..352
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 355..359
FT /evidence="ECO:0007829|PDB:4AR9"
FT TURN 360..363
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 364..368
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 374..395
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 401..404
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 406..408
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 409..417
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 418..421
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 424..427
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 433..439
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 440..442
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 444..448
FT /evidence="ECO:0007829|PDB:4AR9"
FT TURN 452..454
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 459..475
FT /evidence="ECO:0007829|PDB:4AR9"
FT TURN 481..483
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 485..487
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 488..490
FT /evidence="ECO:0007829|PDB:4AR9"
FT TURN 491..494
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 495..505
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 517..520
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 527..529
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 533..537
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 541..543
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 547..561
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 563..574
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 578..589
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 592..607
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 609..611
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 619..622
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 630..641
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 647..652
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 654..666
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 673..692
FT /evidence="ECO:0007829|PDB:4AR9"
FT HELIX 697..701
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 703..711
FT /evidence="ECO:0007829|PDB:4AR9"
FT STRAND 717..729
FT /evidence="ECO:0007829|PDB:4AR9"
SQ SEQUENCE 991 AA; 114376 MW; 9A5F57730134DED4 CRC64;
MKKKFIKMLC SIAIGCMIST SYSIKVSAFS NGNTKTNPNG EFKSLSLNST NPYKTKYSFN
DLNKLSNKEI LDLTSKIKWS DISDLFQYNK DSYTFYSNKE RVQALIDGLY EKGCNYTSTD
DKGIDTLVEI LRSGFYLGYY NDSLKYLNDK SFKDKCIPAM IAIENNKNFK LGENGQDTVV
HALGKLIGNT SCNDEVVNKT IPILEQYYNE IDKYSKDRLK SNAVYNFMKE INYDISQYEY
AHNIRDYKNT PWSGKIDSFI DTISKFASIS NVTKDNGWII NNSIYYTAKL SKYHSNPSIP
HSVIDNCIEI FPDYSEQYFT AIEAIKEDFN SRDSKGNVID INKLIEEGKK HYLPKTYTFD
NGKIIIKAGD KVEESKIQKL YWASKEVKSQ FHRIIGNDKP LEVGNADDIL TIVIYNNPEE
YKLNKTLYGY SVDNGGIYIE GIGTFFTYER TPQESIYSLE ELFRHEFTHY LQGRYLIPGL
FNKGDFYKGN NGRITWFEEG SAEFFAGSTR TSVLPRKSMV GGLSKNPKER FNADKLLHSK
YSDGWDFYKY GYAFSDYMYN NNKKLFSDLV STMKNNDVKG YEALIEESSK DSKINKDYEY
HMENLVNNYD NYTIPLVSDD YMKQYDNKSL HEIKSDIEKA MDVKNSQITK ESSQYFDTYN
LKATYTLSSN KGEISNWNYM NNKINEALNK LDNLSWGGYK TVTAYFSNPR LNSNNEVVYD
IVFHGLLSHN KNSNEKVEVK EEPEIKDKDS FENVIYEKEN NDSFDKANKI HKNQIVMATL
DTEDYRDTFY FDALTSGSID ITIENIHGNS DAFNWLVYND EDLNNYIAYP TKKEDNKLMG
SFKVHKPGRY YILVYKTSLN KVNYKLNISD ATNMAPVIKK IHEKENNDSF ETANKITLDT
LVLGNLDYKD VSDIYSFDIE NTKDLNIKLT NLNNLGIAWN LYKESDLNNY IAYGAKSDNA
IVGKCNLSPG KYYLYVYKYS GDKGNYSVII N
