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Protein

Collagenase ColG

Gene

colG

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). Active on soluble type I collagen, insoluble collagen, azocoll, soluble PZ-peptide (all collagenase substrates) and gelatin (PubMed:9922257). The full-length protein has collagenase activity, while the in vivo derived C-terminally truncated shorter versions only act on gelatin (PubMed:9922257). In vitro digestion of soluble calf skin collagen fibrils requires both ColG and ColH; ColG forms missing the second collagen-binding domain are also synergistic with ColH, although their overall efficiency is decreased (PubMed:18374061, PubMed:22099748). The activator domain (residues 119-388) and catalytic subdomain (389-670) open and close around substrate using a Gly-rich hinge (387-397), allowing digestion when the protein is closed (PubMed:21947205, PubMed:23703618). Binding of collagen requires Ca2+ and is inhibited by EGTA; the collagen-binding domain (CBD, S3a plus S3b) specifically recognizes the triple-helical conformation made by 3 collagen protein chains in the triple-helical region (PubMed:11121400). Isolated CBD (S3a plus S3b) binds collagen fibrils and sheets of many tissues (PubMed:11913772).11 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'.1 Publication2 Publications

Cofactori

Protein has several cofactor binding sites:

Enzyme regulationi

Inhibited by 1-10-phenanthroline (PubMed:18937627). Inhibited by peptidomimetic isoamyl-phosphonyl-Gly-Pro-Ala, which binds to Zn2+ (PubMed:21947205). 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/sec, using a catalytic fragment (119-790) on an artificial substrate.1 Publication
  1. KM=0.840 mM for furylacryloyl-Leu-Gly-Pro-Ala (FALGPA)1 Publication
  1. Vmax=0.0852 µmol/min/mg enzyme1 Publication

Sites

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Metal bindingi498Calcium 11 PublicationBy similarity1
Metal bindingi523Zinc; catalytic; via tele nitrogenPROSITE-ProRule annotationCombined sources2 Publications1
Active sitei524PROSITE-ProRule annotation1 Publication1
Metal bindingi527Zinc; catalytic; via tele nitrogenPROSITE-ProRule annotationCombined sources2 Publications1
Metal bindingi531Calcium 1; via carbonyl oxygen1 PublicationBy similarity1
Metal bindingi535Calcium 1; via carbonyl oxygen1 PublicationBy similarity1
Metal bindingi537Calcium 1; via carbonyl oxygen1 PublicationBy similarity1
Metal bindingi555Zinc; catalyticCombined sources2 Publications1
Metal bindingi795Calcium 2Combined sources1 Publication1
Metal bindingi796Calcium 2; via carbonyl oxygenCombined sources1 Publication1
Metal bindingi823Calcium 2Combined sources1 Publication1
Metal bindingi825Calcium 2Combined sources1 Publication1
Metal bindingi864Calcium 2Combined sources1 Publication1
Metal bindingi890Calcium 3Combined sources1
Metal bindingi892Calcium 3Combined sources1
Metal bindingi892Calcium 4Combined sources1
Metal bindingi894Calcium 4Combined sources1
Metal bindingi913Calcium 4; via carbonyl oxygenCombined sources1
Metal bindingi918Calcium 3Combined sources1
Metal bindingi918Calcium 4Combined sources1
Metal bindingi920Calcium 4; via carbonyl oxygenCombined sources1
Metal bindingi921Calcium 3Combined sources1
Metal bindingi921Calcium 4Combined sources1
Sitei1006C-terminus of form C1b1 Publication1
Metal bindingi1009Calcium 5Combined sources2 Publications1
Metal bindingi1011Calcium 5Combined sources2 Publications1
Metal bindingi1011Calcium 6Combined sources2 Publications1
Metal bindingi1013Calcium 6Combined sources2 Publications1
Metal bindingi1014Calcium 6Combined sources2 Publications1
Sitei1018C-terminus of form C1c1 Publication1
Metal bindingi1032Calcium 5; via carbonyl oxygenCombined sources2 Publications1
Metal bindingi1037Calcium 5Combined sources2 Publications1
Metal bindingi1037Calcium 6Combined sources2 Publications1
Metal bindingi1039Calcium 6; via carbonyl oxygenCombined sources2 Publications1
Metal bindingi1040Calcium 5Combined sources2 Publications1
Metal bindingi1040Calcium 6Combined sources2 Publications1
Binding sitei1067Collagen-binding2 Publications1
Binding sitei1080Collagen-binding2 Publications1

GO - Molecular functioni

  • calcium ion binding Source: UniProtKB
  • collagen binding Source: UniProtKB
  • endopeptidase activity Source: UniProtKB
  • metalloendopeptidase activity Source: UniProtKB
  • serine-type endopeptidase activity Source: InterPro
  • tripeptidase activity Source: UniProtKB
  • zinc ion binding Source: UniProtKB

Keywordsi

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

Enzyme and pathway databases

BRENDAi3.4.24.3. 1481.

Protein family/group databases

MEROPSiM09.002.

Names & Taxonomyi

Protein namesi
Recommended name:
Collagenase ColG1 Publication (EC:3.4.24.31 Publication)
Alternative name(s):
Class I collagenase2 Publications
Gelatinase ColG1 Publication
Microbial collagenase
Gene namesi
Name:colG1 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 their 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
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 reduce these joint contractures (PubMed:10913202, PubMed:19726771). 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 psychosocial symptoms (PubMed:8417217, PubMed:25711400).4 Publications

Mutagenesis

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Mutagenesisi389 – 397Missing : Degrades soluble FALGPA peptide (furylacryloyl-Leu-Gly-Pro-Ala) but only 40% active on type I collagen. 1 Publication9
Mutagenesisi524E → D: Retains 4% digestion of collagen, still bind collagen. 1 Publication1
Mutagenesisi1039R → A: S3b fragment has 2.3-fold increased affinity for collagen. 1 Publication1
Mutagenesisi1062F → A: S3b fragment has 2.4-fold increased affinity for collagen. 1 Publication1
Mutagenesisi1067T → A: S3b fragment has 10-fold decreased affinity for collagen. 1 Publication1
Mutagenesisi1080Y → A: S3b fragment has 12-fold decreased affinity for collagen. 1 Publication1
Mutagenesisi1088V → A: S3b fragment has 2.2-fold increased affinity for collagen. 1 Publication1
Mutagenesisi1102L → A: S3b fragment has 5-fold decreased affinity for collagen. 1 Publication1
Mutagenesisi1104Y → A or S: S3b fragment has no collagen binding. 1 Publication1
Mutagenesisi1104Y → F: S3b fragment has 12-fold decreased affinity for collagen. 1 Publication1
Mutagenesisi1106Y → A: S3b fragment has 40-fold decreased affinity for collagen. 1 Publication1

Protein family/group databases

Allergomei5989. Clo hi Collagenase.

Chemistry databases

ChEMBLiCHEMBL2268009.

PTM / Processingi

Molecule processing

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Signal peptidei1 – 45Sequence analysisAdd BLAST45
PropeptideiPRO_000044354546 – 1101 PublicationAdd BLAST65
ChainiPRO_0000443546111 – 1118Collagenase ColGAdd BLAST1008

Post-translational modificationi

Upon purification gives 67 kDa, 78 kDa, 82 kDa and 116 kDa (full-length) proteins all of which have the same N-terminus; only the longest form digests insoluble collagen (PubMed:9922257). At least 2 in vivo isolated forms (C1b and C1c) are missing the second collagen-binding domain, ending on Lys-1006 and Lys-1018 respectively (PubMed:22099748).2 Publications

Keywords - PTMi

Zymogen

Proteomic databases

PRIDEiQ9X721.

Expressioni

Inductioni

RNA levels are high in late logarithmic phase.1 Publication

Interactioni

GO - Molecular functioni

  • collagen binding Source: UniProtKB

Chemistry databases

BindingDBiQ9S0X0.

Structurei

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
PDB entryMethodResolution (Å)ChainPositionsPDBsum
1NQDX-ray1.65A/B1003-1118[»]
1NQJX-ray1.00A/B1003-1118[»]
2O8OX-ray1.35A/B1003-1118[»]
2Y3UX-ray2.55A119-880[»]
2Y50X-ray2.80A119-880[»]
2Y6IX-ray3.25A119-880[»]
2Y72X-ray1.18A/B799-880[»]
4AQOX-ray0.99A792-880[»]
4AREX-ray2.19A119-789[»]
4HPKX-ray1.35A1006-1118[»]
B1003-1118[»]
4JRWX-ray1.60A/B797-881[»]
4TN9X-ray1.40A/B797-881[»]
5IKUX-ray1.90A883-1118[»]
ProteinModelPortaliQ9X721.
SMRiQ9X721.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiQ9X721.

Family & Domainsi

Domains and Repeats

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Domaini797 – 885PKDPROSITE-ProRule annotationAdd BLAST89

Region

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Regioni111 – 786S1 metalloprotease domain, degrades both FALGPA (furylacryloyl-Leu-Gly-Pro-Ala) and type I collagen2 Publications1 PublicationAdd BLAST676
Regioni119 – 388Activator domain required for full activity on collagen1 Publication1 PublicationAdd BLAST270
Regioni389 – 670Catalytic subdomain1 PublicationAdd BLAST282
Regioni396 – 1118Degrades soluble FALGPA peptide (furylacryloyl-Leu-Gly-Pro-Ala) but not type I collagen1 PublicationAdd BLAST723
Regioni679 – 790Helper subdomain1 PublicationAdd BLAST112
Regioni787 – 882S2 domain2 PublicationsAdd BLAST96
Regioni886 – 1003S3a collagen-binding domain2 PublicationsAdd BLAST118
Regioni1008 – 1118S3b collagen-binding domain2 Publications1 PublicationAdd BLAST111
Regioni1102 – 1106Collagen-binding2 Publications5

Domaini

The mature protein has 4 domains; a metalloprotease domain (S1, approximately residues 111-786), S2 (877-882, equivalent to PKD), and 2 collagen-binding domains (CBD) S3a (997-1003) and S3b (1008-1118) (PubMed:9922257, PubMed:11121400). The S1 domain has collagen hydrolytic activity (PubMed:11121400, PubMed:18937627). The metalloprotease S1 domain is composed of 3 subdomains which together resemble a saddle; an activator domain (residues 119-388), the catalytic peptidase subdomain (398-670) and a helper subdomain (679-790) joined by a Gly-rich hinge (387-397) (PubMed:21947205, PubMed:23703618). The S2 domain (799-880, PKD) is flexible within a larger structure (S1 plus S2, residues 119-880) (PubMed:21947205, PubMed:21871007). Binding to Ca2+ renders the midsection of S2 more flexible; Ca2+ binding confers thermostability (PubMed:25760606). S3a and S3b each have collagen-binding activity; collagen is bound more efficiently when both S3a and S3b are present (PubMed:11121400). CBD S3a plus S3b binds to many types of collagen in vitro and in vivo (PubMed:11913772). The structure of CBD S3b becomes more compact and thermostable when it is bound to Ca2+ and its N-terminal linker (approximately residues 1008-1020) changes from an extended alpha-helix to a beta-sheet anchored to the rest of the CBD (PubMed:12682007, PubMed:23144249). S3b may act as a Ca2+-activated molecular switch to trigger domain reorientation (PubMed:12682007). Isolated CBD S3b binds unidirectionally to the C-terminus of the collagen triple helix via a surface cleft (PubMed:19208618, PubMed:23144249). The S3b domain binds preferentially to undertwisted segions of collagen (PubMed:22898990).4 Publications10 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. 1 hit.
InterProiView protein in InterPro
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. 1 hit.
PRINTSiPR00931. MICOLLPTASE.
SMARTiView protein in SMART
SM00089. PKD. 1 hit.
SUPFAMiSSF49299. SSF49299. 1 hit.
PROSITEiView protein in PROSITE
PS50093. PKD. 1 hit.
PS00142. ZINC_PROTEASE. 1 hit.

Sequencei

Sequence statusi: Complete.

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

Q9X721-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MKKNILKILM DSYSKESKIQ TVRRVTSVSL LAVYLTMNTS SLVLAKPIEN
60 70 80 90 100
TNDTSIKNVE KLRNAPNEEN SKKVEDSKND KVEHVKNIEE AKVEQVAPEV
110 120 130 140 150
KSKSTLRSAS IANTNSEKYD FEYLNGLSYT ELTNLIKNIK WNQINGLFNY
160 170 180 190 200
STGSQKFFGD KNRVQAIINA LQESGRTYTA NDMKGIETFT EVLRAGFYLG
210 220 230 240 250
YYNDGLSYLN DRNFQDKCIP AMIAIQKNPN FKLGTAVQDE VITSLGKLIG
260 270 280 290 300
NASANAEVVN NCVPVLKQFR ENLNQYAPDY VKGTAVNELI KGIEFDFSGA
310 320 330 340 350
AYEKDVKTMP WYGKIDPFIN ELKALGLYGN ITSATEWASD VGIYYLSKFG
360 370 380 390 400
LYSTNRNDIV QSLEKAVDMY KYGKIAFVAM ERITWDYDGI GSNGKKVDHD
410 420 430 440 450
KFLDDAEKHY LPKTYTFDNG TFIIRAGDKV SEEKIKRLYW ASREVKSQFH
460 470 480 490 500
RVVGNDKALE VGNADDVLTM KIFNSPEEYK FNTNINGVST DNGGLYIEPR
510 520 530 540 550
GTFYTYERTP QQSIFSLEEL FRHEYTHYLQ ARYLVDGLWG QGPFYEKNRL
560 570 580 590 600
TWFDEGTAEF FAGSTRTSGV LPRKSILGYL AKDKVDHRYS LKKTLNSGYD
610 620 630 640 650
DSDWMFYNYG FAVAHYLYEK DMPTFIKMNK AILNTDVKSY DEIIKKLSDD
660 670 680 690 700
ANKNTEYQNH IQELADKYQG AGIPLVSDDY LKDHGYKKAS EVYSEISKAA
710 720 730 740 750
SLTNTSVTAE KSQYFNTFTL RGTYTGETSK GEFKDWDEMS KKLDGTLESL
760 770 780 790 800
AKNSWSGYKT LTAYFTNYRV TSDNKVQYDV VFHGVLTDNA DISNNKAPIA
810 820 830 840 850
KVTGPSTGAV GRNIEFSGKD SKDEDGKIVS YDWDFGDGAT SRGKNSVHAY
860 870 880 890 900
KKAGTYNVTL KVTDDKGATA TESFTIEIKN EDTTTPITKE MEPNDDIKEA
910 920 930 940 950
NGPIVEGVTV KGDLNGSDDA DTFYFDVKED GDVTIELPYS GSSNFTWLVY
960 970 980 990 1000
KEGDDQNHIA SGIDKNNSKV GTFKSTKGRH YVFIYKHDSA SNISYSLNIK
1010 1020 1030 1040 1050
GLGNEKLKEK ENNDSSDKAT VIPNFNTTMQ GSLLGDDSRD YYSFEVKEEG
1060 1070 1080 1090 1100
EVNIELDKKD EFGVTWTLHP ESNINDRITY GQVDGNKVSN KVKLRPGKYY
1110
LLVYKYSGSG NYELRVNK
Length:1,118
Mass (Da):126,242
Last modified:November 1, 1999 - v1
Checksum:i770CF3F60E4D4FD4
GO

Experimental Info

Feature keyPosition(s)DescriptionActionsGraphical viewLength
Sequence conflicti606F → V in BAA86030 (Ref. 2) Curated1
Sequence conflicti656 – 659EYQN → DYQT in BAA86030 (Ref. 2) Curated4
Sequence conflicti836 – 837GD → VY in BAA86030 (Ref. 2) Curated2

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
D87215 Genomic DNA. Translation: BAA77453.1.
AB026889 Genomic DNA. Translation: BAA86030.1.

Entry informationi

Entry nameiCOLG_HATHI
AccessioniPrimary (citable) accession number: Q9X721
Secondary accession number(s): Q9S0X0
Entry historyiIntegrated into UniProtKB/Swiss-Prot: March 28, 2018
Last sequence update: November 1, 1999
Last modified: March 28, 2018
This is version 90 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