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Protein

Collagenase ColH

Gene

colH

Organism
Hathewaya histolytica (Clostridium histolyticum)
Status
Reviewed-Annotation score: -Experimental evidence at protein leveli

Functioni

Clostridial collagenases are among the most efficient degraders of eukaryotic collagen known; saprophytes use collagen as a carbon source while pathogens additionally digest collagen to aid in host colonization. Has both tripeptidylcarboxypeptidase on Gly-X-Y and endopeptidase activities; the endopeptidase cuts within the triple helix region of collagen while tripeptidylcarboxypeptidase successively digests the exposed ends, thus clostridial collagenases can digest large sections of collagen (PubMed:3002446). The full-length protein has collagenase activity, while both the 116 kDa and 98 kDa forms act on gelatin (PubMed:7961400). In vitro digestion of soluble calf skin collagen fibrils requires both ColG and ColH; ColG forms missing the second collagen-binding domain is also synergistic with ColH, although their overall efficiency is decreased (PubMed:18374061, PubMed:22099748). Digestion of collagen requires Ca2+ and is inhibited by EDTA (PubMed:9452493). The activator domain (residues 119-388) and catalytic subdomain (330-601) open and close around substrate allowing digestion when the protein is closed (PubMed:23703618).9 Publications

Miscellaneous

Clostridial collagenases enable the bacteria to infiltrate and colonize host tissue, and contribute to gas gangrene (myonecrosis) pathogenesis.Curated

Catalytic activityi

Digestion of native collagen in the triple helical region at Xaa-|-Gly bonds. With synthetic peptides, a preference is shown for Gly at P3 and P1'; Pro and Ala at P2 and P2'; and hydroxyproline, Ala or Arg at P3'.3 Publications2 Publications

Cofactori

Protein has several cofactor binding sites:

Enzyme regulationi

Inhibited by EDTA (PubMed:9452493). Inhibited by 1-10-phenanthroline (PubMed:18937627). Inhibited by broad-spectrum zinc metalloprotease inhibitor batimastat (PubMed:28820255). N-aryl mercaptoacetamide-based inhibitors have been isolated that act on clostridial collagenases with submicromolar affinity while having negligibile activity on human collagenases (PubMed:28820255).3 Publications

Kineticsi

kcat is 0.11 per second on Pz peptide with whole enzyme (PubMed:10217773). kcat is 15.9 per second on FALGPA with a catalytic fragment (residues 41-717) (PubMed:18937627).2 Publications
  1. KM=0.88 mM for Pz peptide (4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg)1 Publication
  2. KM=0.269 mM for furylacryloyl-Leu-Gly-Pro-Ala (FALGPA) with a catalytic fragment (residues 41-717)1 Publication
  3. KM=62 µM for (7-Methoxycoumarin-4-yl)acetyl-Ala-Gly-Pro-Pro-Gly-Pro-N-3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl)-Gly-Arg-NH2 with peptidase fragment (residues 331-721)1 Publication
  1. Vmax=12.1 µmol/min/mg enzyme on FALGPA with a catalytic fragment (residues 41-717)1 Publication

Sites

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Metal bindingi421Zinc; catalyticCombined sources1 Publication1
Metal bindingi430Calcium 1Combined sources2 Publications1
Metal bindingi455Zinc; catalytic; via tele nitrogenPROSITE-ProRule annotationCombined sources1 Publication2 Publications1
Active sitei456PROSITE-ProRule annotation1 Publication1
Metal bindingi459Zinc; catalytic; via tele nitrogenPROSITE-ProRule annotationCombined sources1 Publication2 Publications1
Metal bindingi463Calcium 1; via carbonyl oxygenCombined sources2 Publications1
Metal bindingi467Calcium 1; via carbonyl oxygenCombined sources2 Publications1
Metal bindingi469Calcium 1; via carbonyl oxygenCombined sources2 Publications1
Metal bindingi487Zinc; catalyticCombined sources1 Publication2 Publications1
Metal bindingi725Calcium 2Combined sources1 Publication1
Metal bindingi726Calcium 2; via carbonyl oxygenCombined sources1 Publication1
Metal bindingi753Calcium 2Combined sources1 Publication1
Metal bindingi755Calcium 2Combined sources1 Publication1
Metal bindingi794Calcium 2Combined sources1 Publication1
Metal bindingi814Calcium 3Combined sources1 Publication1
Metal bindingi815Calcium 3; via carbonyl oxygenCombined sources1 Publication1
Metal bindingi842Calcium 3Combined sources1 Publication1
Metal bindingi844Calcium 3Combined sources1 Publication1
Metal bindingi884Calcium 3Combined sources1 Publication1
Metal bindingi908Calcium 4Combined sources1 Publication1
Metal bindingi910Calcium 4Combined sources1 Publication1
Metal bindingi910Calcium 5Combined sources1 Publication1
Metal bindingi912Calcium 5Combined sources1 Publication1
Metal bindingi913Calcium 5Combined sources1 Publication1
Metal bindingi931Calcium 4; via carbonyl oxygenCombined sources1 Publication1
Metal bindingi937Calcium 4Combined sources1 Publication1
Metal bindingi937Calcium 5Combined sources1 Publication1
Metal bindingi938Calcium 5; via carbonyl oxygenCombined sources1 Publication1
Metal bindingi939Calcium 4Combined sources1 Publication1
Metal bindingi939Calcium 5Combined sources1 Publication1
Binding sitei977Collagen-binding1 PublicationBy similarity1

GO - Molecular functioni

  • calcium ion binding Source: UniProtKB
  • collagen binding Source: UniProtKB
  • endopeptidase activity Source: UniProtKB
  • metalloendopeptidase activity Source: UniProtKB
  • tripeptidase activity Source: UniProtKB
  • zinc ion binding Source: UniProtKB

GO - Biological processi

  • collagen metabolic process Source: UniProtKB
  • pathogenesis Source: UniProtKB-KW

Keywordsi

Molecular functionHydrolase, Metalloprotease, Protease
Biological processVirulence
LigandCalcium, Metal-binding, Zinc

Protein family/group databases

MEROPSiM09.003

Names & Taxonomyi

Protein namesi
Recommended name:
Collagenase ColH1 Publication (EC:3.4.24.31 Publication)
Alternative name(s):
Class II collagenase2 Publications
Gelatinase ColH1 Publication
Microbial collagenase
Gene namesi
Name:colH1 Publication
OrganismiHathewaya histolytica (Clostridium histolyticum)
Taxonomic identifieri1498 [NCBI]
Taxonomic lineageiBacteriaFirmicutesClostridiaClostridialesClostridiaceaeHathewaya

Subcellular locationi

GO - Cellular componenti

  • extracellular region Source: UniProtKB

Keywords - Cellular componenti

Secreted

Pathology & Biotechi

Biotechnological usei

Widely used for tissue dissociation due to its potent activity on connective tissue.Curated
A mix of ColG and ColH is used for isolation of pancreatic islet cells for subsequent transplantation.2 Publications
A mix of ColG and ColH has been used to allow release of retained placenta in cows and mares, and its use in humans has been proposed.1 Publication
Has been used to anchor otherwise diffusible proteins to the host extracellular matrix. Fusions between the collagen-binding domain (S2B plus S3) and rat epidermal growth factor or human basic fibroblast growth factor (bFGF) were injected subcutaneously into adult male BALB/c-nu mice. The fusion proteins remained at the sites of injection for up to 10 days, and bFGF strongly stimulated fibroblast growth (PubMed:9618531).1 Publication
N-aryl mercaptoacetamide-based inhibitors with submicromolar affinity for clostridial collagenases but negligibile activity on human collagenases have been discovered that may lead to promising anti-infective drugs against Clostridia (PubMed:28820255).1 Publication

Pharmaceutical usei

SANTYL Ointment (Smith and Nephew, Inc.) is indicated for debriding chronic dermal ulcers and severely burned areas. It is unclear which of the collagenases from this bacteria is in the ointment.1 Publication
Xiaflex (Endo Pharmaceuticals, Inc.) is a mix of H.histolytica collagenases (ColG and ColH) used to treat both Dupuytren disease and Peyronie disease. Dupuytren disease is a progressive genetic disorder of pathologic collagen production and deposition under the skin of the hand that causes the fingers to be drawn into the palm, leading to flexion contractures of the joints, which can severely limit hand function. Injections of collagenase may reduce these joint contractures (PubMed:19726771, PubMed:10913202). Peyronie disease (PD) is characterized by a disorganized, excessive deposition of collagen that forms a plaque within the penis. The plaque restricts lengthening on the affected side during erection, which can lead to penile curvature deformity, discomfort and erectile dysfunction, and can eventually lead to psychosocial effects such as depression and relationship difficulties. Studies have shown the clinical efficacy of collagenase injection for reducing penile curvature deformity and PD symptom bother (PubMed:8417217, PubMed:25711400).4 Publications

Mutagenesis

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Mutagenesisi426G → V: Loss of activity, in a catalytic fragment (residues 41-717) using FALGPA as substrate. 1 Publication1
Mutagenesisi455H → A: No collagen degradation, about 50% zinc content. 1 Publication1
Mutagenesisi455H → F: No collagen degradation, about 10% zinc content. 1 Publication1
Mutagenesisi456E → D or Q: No collagen degradation, wild-type zinc content. 1 Publication1
Mutagenesisi456E → D: Does not degrade collagen, still binds collagen. 1 Publication1
Mutagenesisi459H → R: No collagen degradation, about 20% zinc content. 1 Publication1
Mutagenesisi479N → A: Wild-type collagen degradation and zinc content. 1 Publication1
Mutagenesisi486E → A or Q: About 15% collagen degradation, wild-type zinc content. KM for PZ peptide is wild-type, kcat decreases 4-fold for Ala-486. 1 Publication1
Mutagenesisi486E → D: About 50% collagen degradation, wild-type zinc content. 1 Publication1
Mutagenesisi487E → A, D or Q: Less than 5% collagen degradation, 20-42% zinc content. KM for PZ peptide is 75%, kcat decreases 20-fold for Gln-487. 1 Publication1
Mutagenesisi487E → H: Slight decrease in inhibition of collagenase activity by an N-aryl mercaptoacetamide-based inhibitor. 1 Publication1
Mutagenesisi491E → A, D or Q: 5 to 12% collagen degradation, 70% to wild-type zinc content. KM for PZ peptide is nearly wild-type, kcat decreases 15-fold for Ala-491. 1 Publication1

PTM / Processingi

Molecule processing

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Signal peptidei1 – 30Sequence analysisAdd BLAST30
PropeptideiPRO_000044354731 – 401 Publication10
ChainiPRO_501084360541 – 1021Collagenase ColHAdd BLAST981

Post-translational modificationi

Upon purification gives rise to 98 kDa, 105 kDa and 116 kDa (full-length) proteins, all of which have the same N-terminus (PubMed:7961400, PubMed:9922257).2 Publications

Keywords - PTMi

Zymogen

Structurei

Secondary structure

11021
Legend: HelixTurnBeta strandPDB Structure known for this area
Show more details
Feature keyPosition(s)DescriptionActionsGraphical viewLength
Beta strandi345 – 349Combined sources5
Turni350 – 353Combined sources4
Beta strandi354 – 358Combined sources5
Helixi364 – 385Combined sources22
Beta strandi391 – 394Combined sources4
Helixi396 – 398Combined sources3
Beta strandi399 – 407Combined sources9
Helixi408 – 414Combined sources7
Helixi415 – 418Combined sources4
Beta strandi423 – 429Combined sources7
Helixi430 – 432Combined sources3
Beta strandi434 – 438Combined sources5
Helixi442 – 444Combined sources3
Helixi449 – 465Combined sources17
Helixi475 – 480Combined sources6
Helixi483 – 493Combined sources11
Beta strandi498 – 500Combined sources3
Helixi506 – 509Combined sources4
Turni510 – 514Combined sources5
Helixi517 – 519Combined sources3
Helixi523 – 526Combined sources4
Helixi536 – 551Combined sources16
Helixi553 – 564Combined sources12
Helixi568 – 579Combined sources12
Helixi582 – 597Combined sources16
Helixi599 – 601Combined sources3
Helixi609 – 612Combined sources4
Helixi620 – 630Combined sources11
Beta strandi634 – 642Combined sources9
Beta strandi647 – 659Combined sources13
Helixi663 – 681Combined sources19
Helixi687 – 691Combined sources5
Beta strandi693 – 701Combined sources9
Beta strandi705 – 717Combined sources13
Beta strandi736 – 739Combined sources4
Beta strandi743 – 745Combined sources3
Beta strandi758 – 764Combined sources7
Beta strandi766 – 768Combined sources3
Beta strandi777 – 779Combined sources3
Beta strandi784 – 794Combined sources11
Beta strandi799 – 809Combined sources11
Helixi811 – 813Combined sources3
Beta strandi825 – 828Combined sources4
Beta strandi831 – 834Combined sources4
Beta strandi847 – 853Combined sources7
Beta strandi855 – 857Combined sources3
Beta strandi867 – 869Combined sources3
Beta strandi874 – 884Combined sources11
Beta strandi889 – 899Combined sources11
Helixi915 – 917Combined sources3
Beta strandi928 – 932Combined sources5
Beta strandi939 – 947Combined sources9
Beta strandi949 – 957Combined sources9
Beta strandi959 – 968Combined sources10
Beta strandi974 – 977Combined sources4
Beta strandi982 – 991Combined sources10
Beta strandi995 – 1005Combined sources11
Beta strandi1009 – 1017Combined sources9

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
PDB entryMethodResolution (Å)ChainPositionsPDBsum
3JQWX-ray2.00A/B/C902-1021[»]
3JQXX-ray2.20A/B/C902-1021[»]
4AR1X-ray2.01A331-721[»]
4ARFX-ray1.77A331-721[»]
4JGUX-ray1.42A/B806-900[»]
4U6TX-ray1.76A/B/C/D725-810[»]
4U7KX-ray1.91A/B/C/D/E/F/G/H724-810[»]
5O7EX-ray1.87A331-721[»]
SMRiQ46085
ModBaseiSearch...
MobiDBiSearch...

Family & Domainsi

Domains and Repeats

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Domaini727 – 808PKD 1PROSITE-ProRule annotationAdd BLAST82
Domaini816 – 905PKD 2PROSITE-ProRule annotationAdd BLAST90

Region

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Regioni41 – 717S1 metalloprotease domain2 PublicationsAdd BLAST677
Regioni41 – 320Activator domain1 PublicationAdd BLAST280
Regioni330 – 601Catalytic subdomain1 PublicationAdd BLAST272
Regioni609 – 721Helper subdomain1 PublicationAdd BLAST113
Regioni718 – 810S2a domain2 PublicationsAdd BLAST93
Regioni811 – 904S2b domain2 PublicationsAdd BLAST94
Regioni905 – 1021S3 collagen-binding domain2 PublicationsAdd BLAST117
Regioni1002 – 1004Collagen-binding1 PublicationBy similarity3

Domaini

The mature protein has 4 domains; a metalloprotease domain (S1, approximately residues 41-717), S2a (718-810, equivalent to PKD 1), S2b (811-904, equivalent to PKD 2) and a collagen-binding domain (S3, 905-1021) (PubMed:9922257, PubMed:9452493). The protein has an elongated shape, which lengthens further in calcium-chelated conditions; calcium-chelation also increases its susceptibility to exogenous proteases (PubMed:23561530). The S1 domain has the collagen hydrolytic activity (PubMed:9452493, PubMed:18937627). The metalloprotease S1 domain is composed of 3 subdomains which together resemble a saddle; an activator domain (residues 41-320), the catalytic peptidase subdomain (330-601) and a helper subdomain (609-721) (PubMed:23703618). Unlike the S2 domain in ColG, upon binding to Ca2+ the midsections of S2a and S2b remain rigid; Ca2+ binding confers thermostability (PubMed:25760606). Ca2+-binding also alters the orientation of the N-terminal linker of S2b so it lies along the long axis of the domain (PubMed:25760606). S3 has collagen-binding activity (PubMed:9452493). The structure of S3 becomes more thermostable when it is bound to Ca2+ (PubMed:23144249). Isolated CBD S3 binds unidirectionally to the C-terminus of the collagen triple helix via a surface cleft (PubMed:23144249).3 Publications6 Publications

Sequence similaritiesi

Belongs to the peptidase M9B family. Collagenase subfamily.2 Publications

Keywords - Domaini

Repeat, Signal

Family and domain databases

Gene3Di2.60.40.10, 2 hits
InterProiView protein in InterPro
IPR013320 ConA-like_dom_sf
IPR013783 Ig-like_fold
IPR013661 Peptidase_M9_N_dom
IPR002169 Peptidase_M9A/M9B
IPR022409 PKD/Chitinase_dom
IPR000601 PKD_dom
IPR035986 PKD_dom_sf
PfamiView protein in Pfam
PF01752 Peptidase_M9, 1 hit
PF08453 Peptidase_M9_N, 1 hit
PF00801 PKD, 2 hits
PRINTSiPR00931 MICOLLPTASE
SMARTiView protein in SMART
SM00089 PKD, 2 hits
SUPFAMiSSF49299 SSF49299, 2 hits
SSF49899 SSF49899, 1 hit
PROSITEiView protein in PROSITE
PS50093 PKD, 2 hits
PS00142 ZINC_PROTEASE, 1 hit

Sequencei

Sequence statusi: Complete.

Sequence processingi: The displayed sequence is further processed into a mature form.

Q46085-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MKRKCLSKRL MLAITMATIF TVNSTLPIYA AVDKNNATAA VQNESKRYTV
60 70 80 90 100
SYLKTLNYYD LVDLLVKTEI ENLPDLFQYS SDAKEFYGNK TRMSFIMDEI
110 120 130 140 150
GRRAPQYTEI DHKGIPTLVE VVRAGFYLGF HNKELNEINK RSFKERVIPS
160 170 180 190 200
ILAIQKNPNF KLGTEVQDKI VSATGLLAGN ETAPPEVVNN FTPILQDCIK
210 220 230 240 250
NIDRYALDDL KSKALFNVLA APTYDITEYL RATKEKPENT PWYGKIDGFI
260 270 280 290 300
NELKKLALYG KINDNNSWII DNGIYHIAPL GKLHSNNKIG IETLTEVMKV
310 320 330 340 350
YPYLSMQHLQ SADQIKRHYD SKDAEGNKIP LDKFKKEGKE KYCPKTYTFD
360 370 380 390 400
DGKVIIKAGA RVEEEKVKRL YWASKEVNSQ FFRVYGIDKP LEEGNPDDIL
410 420 430 440 450
TMVIYNSPEE YKLNSVLYGY DTNNGGMYIE PEGTFFTYER EAQESTYTLE
460 470 480 490 500
ELFRHEYTHY LQGRYAVPGQ WGRTKLYDND RLTWYEEGGA ELFAGSTRTS
510 520 530 540 550
GILPRKSIVS NIHNTTRNNR YKLSDTVHSK YGASFEFYNY ACMFMDYMYN
560 570 580 590 600
KDMGILNKLN DLAKNNDVDG YDNYIRDLSS NYALNDKYQD HMQERIDNYE
610 620 630 640 650
NLTVPFVADD YLVRHAYKNP NEIYSEISEV AKLKDAKSEV KKSQYFSTFT
660 670 680 690 700
LRGSYTGGAS KGKLEDQKAM NKFIDDSLKK LDTYSWSGYK TLTAYFTNYK
710 720 730 740 750
VDSSNRVTYD VVFHGYLPNE GDSKNSLPYG KINGTYKGTE KEKIKFSSEG
760 770 780 790 800
SFDPDGKIVS YEWDFGDGNK SNEENPEHSY DKVGTYTVKL KVTDDKGESS
810 820 830 840 850
VSTTTAEIKD LSENKLPVIY MHVPKSGALN QKVVFYGKGT YDPDGSIAGY
860 870 880 890 900
QWDFGDGSDF SSEQNPSHVY TKKGEYTVTL RVMDSSGQMS EKTMKIKITD
910 920 930 940 950
PVYPIGTEKE PNNSKETASG PIVPGIPVSG TIENTSDQDY FYFDVITPGE
960 970 980 990 1000
VKIDINKLGY GGATWVVYDE NNNAVSYATD DGQNLSGKFK ADKPGRYYIH
1010 1020
LYMFNGSYMP YRINIEGSVG R
Length:1,021
Mass (Da):116,377
Last modified:November 1, 1996 - v1
Checksum:i826F45EFD96F917C
GO

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AB014075 Genomic DNA Translation: BAA34542.1
D29981 Genomic DNA Translation: BAA06251.1
PIRiI40805

Entry informationi

Entry nameiCOLH_HATHI
AccessioniPrimary (citable) accession number: Q46085
Entry historyiIntegrated into UniProtKB/Swiss-Prot: March 28, 2018
Last sequence update: November 1, 1996
Last modified: May 23, 2018
This is version 115 of the entry and version 1 of the sequence. See complete history.
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Keywords - Technical termi

3D-structure, Direct protein sequencing, Pharmaceutical

Documents

  1. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  2. Peptidase families
    Classification of peptidase families and list of entries
  3. SIMILARITY comments
    Index of protein domains and families

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