Biosynthetic processing of the pro-alpha 1(V)2pro-alpha 2(V) collagen heterotrimer by bone morphogenetic protein-1 and furin-like proprotein convertases.
The low abundance fibrillar collagen type V is incorporated into and regulates the diameters of type I collagen fibrils. Bone morphogenetic protein-1 (BMP-1) is a metalloprotease that plays key roles in regulating formation of vertebrate extracellular matrix; it cleaves the C-propeptides of the major fibrillar procollagens I-III and processes precursors to produce the mature forms of the cross-linking enzyme prolysyl oxidase, the proteoglycan biglycan, and the basement membrane protein laminin 5. Here we have successfully produced recombinant pro-alpha1(V)(2)pro-alpha2(V) heterotrimers, and we have used these to characterize biosynthetic processing of the most prevalent in vivo form of type V procollagen. In addition, we have compared the processing of endogenous pro-alpha1(V) chains by wild type mouse embryo fibroblasts and by fibroblasts derived from embryos doubly homozygous null for the Bmp-1 gene and for a gene encoding the closely related metalloprotease mammalian Tolloid-like 1. Together, results presented herein indicate that within pro-alpha1(V)(2)pro-alpha2(V) heterotrimers, pro-alpha1(V) N-propeptides and pro-alpha2(V) C-propeptides are processed by BMP-1-like enzymes, and pro-alpha1(V) C-propeptides are processed by furin-like proprotein convertases in vivo.
Unsöld
Christine
C
Department of Pathology, University of Wisconsin, Madison, Wisconsin 53706, USA.
Pappano
William N
WN
Imamura
Yasutada
Y
Steiglitz
Barry M
BM
Greenspan
Daniel S
DS
D000595:Amino Acid Sequence; D000818:Animals; D055395:Bone Morphogenetic Protein 1; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002460:Cell Line; D003094:Collagen / Q000096:biosynthesis* / Q000737:chemistry*; D024062:Collagen Type V / Q000096:biosynthesis* / Q000737:chemistry*; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D005347:Fibroblasts / Q000378:metabolism; D045683:Furin; D006801:Humans; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D043484:Proprotein Convertases; D011485:Protein Binding; D011498:Protein Precursors / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D013381:Subtilisins / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
D019485:Bone Morphogenetic Proteins; D024062:Collagen Type V; D018076:DNA, Complementary; D011498:Protein Precursors; D011994:Recombinant Proteins; C426047:pro-alpha1(V-XI) collagen; D003094:Collagen; D043484:Proprotein Convertases; D013381:Subtilisins; D045683:Furin; D008666:Metalloendopeptidases; C525705:BMP1 protein, human; C525707:Bmp1 protein, mouse; D055395:Bone Morphogenetic Protein 1