title: Stepwise proteolytic activation of type I procollagen to collagen within the secretory pathway of tendon fibroblasts in situ.
abstract: Proteolytic cleavage of procollagen I to collagen I is essential for the formation of collagen fibrils in the extracellular matrix of vertebrate tissues. Procollagen is cleaved by the procollagen N- and C-proteinases, which remove the respective N- and C-propeptides from procollagen. Procollagen processing is initiated within the secretory pathway in tendon fibroblasts, which are adept in assembling an ordered extracellular matrix of collagen fibrils in vivo. It was thought that intracellular processing was restricted to the TGN (trans-Golgi network). In the present study, brefeldin A treatment of tendon explant cultures showed that N-proteinase activity is present in the resulting fused ER (endoplasmic reticulum)-Golgi compartment, but that C-proteinase activity is restricted to the TGN in embryonic chick tendon fibroblasts. In late embryonic and postnatal rat tail and postnatal mouse tail tendon, C-proteinase activity was detected in TGN and pre-TGN compartments. Preventing activation of the procollagen N- and C-proteinases with the furin inhibitor Dec-RVKR-CMK (decanoyl-Arg-Val-Lys-Arg-chloromethylketone) indicated that only a fraction of intracellular procollagen cleavage was mediated by newly activated proteinases. In conclusion, the N-propeptides are removed earlier in the secretory pathway than the C-propeptides. The removal of the C-propeptides in post-Golgi compartments most probably indicates preparation of collagen molecules for fibril formation at the cell-matrix interface.
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Leahurst Campus, Chester High Road, Neston CH64 7TE, UK.
authors.forename: Elizabeth G
authors.initials: EG
authors.lastname: Canty-Laird
authors.forename: Yinhui
authors.initials: Y
authors.lastname: Lu
authors.forename: Karl E
authors.initials: KE
authors.lastname: Kadler
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D055395:Bone Morphogenetic Protein 1 / Q000378:metabolism*; D020126:Brefeldin A; D002642:Chick Embryo; D003094:Collagen / Q000096:biosynthesis*; D024042:Collagen Type I / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005347:Fibroblasts / Q000201:enzymology; D051379:Mice; D011348:Procollagen N-Endopeptidase / Q000378:metabolism*; D051381:Rats; D055571:Secretory Pathway / Q000502:physiology*; D013710:Tendons / Q000187:drug effects / Q000196:embryology; D021601:trans-Golgi Network / Q000201:enzymology
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D024042:Collagen Type I; C460143:decanoylRVKRchloromethylketone; D020126:Brefeldin A; D003094:Collagen; D011348:Procollagen N-Endopeptidase; C525707:Bmp1 protein, mouse; D055395:Bone Morphogenetic Protein 1