Mammalian tolloid-like 1 binds procollagen C-proteinase enhancer protein 1 and differs from bone morphogenetic protein 1 in the functional roles of homologous protein domains.
Bone morphogenetic protein 1 (BMP1) is the prototype of a subgroup of metalloproteinases with manifold roles in morphogenesis. Four mammalian subgroup members exist, including BMP1 and mammalian Tolloid-like 1 (mTLL1). Subgroup members have a conserved protein domain structure: an NH2-terminal astacin-like protease domain, followed by a fixed order of CUB and epidermal growth factor-like protein-protein interaction motifs. Previous structure/function studies have documented those BMP1 protein domains necessary for secretion, and activity against various substrates. Here we demonstrate that, in contradiction to previous reports, the most NH2-terminal CUB domain (CUB1) is not required for BMP1 secretion nor is the next CUB domain (CUB2) required for enzymatic activity. The same is true for mTLL1. In fact, secreted protease domains of BMP1 and mTLL1, devoid of CUB or epidermal growth factor-like domains, have procollagen C-proteinase (pCP) activity and activity for biosynthetic processing of biglycan, the latter with kinetics superior to those of the full-length proteins. Structure-function analyses herein also suggest differences in the functional roles played by some of the homologous domains in BMP1 and mTLL1. Surprisingly, although BMP1 has long been known to be Ca2+-dependent, a property previously assumed to apply to all members of the subgroup, mTLL1 is demonstrated to be independent of Ca2 levels in its ability to cleave some, but not all, substrates. We also show that pCP activities of only versions of BMP1 and mTLL1 with intact COOH termini are enhanced by the procollagen C-proteinase enhancer 1 (PCOLCE1) and that mTLL1 binds PCOLCE1, thus suggesting reappraisal of the accepted paradigm for how PCOLCE1 enhances pCP activities.
Ge
Gaoxiang
G
Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison 53706, USA.
Zhang
Yue
Y
Steiglitz
Barry M
BM
Greenspan
Daniel S
DS
D020816:Amino Acid Motifs; D000818:Animals; D015153:Blotting, Western; D055395:Bone Morphogenetic Protein 1; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002918:Chymotrypsin / Q000737:chemistry; D003094:Collagen / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry; D007700:Kinetics; D008666:Metalloendopeptidases / Q000378:metabolism*; D045726:Metalloproteases / Q000737:chemistry / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D013997:Time Factors; D055394:Tolloid-Like Metalloproteinases
D019485:Bone Morphogenetic Proteins; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; C491555:PCOLCE protein, human; D011994:Recombinant Proteins; D003094:Collagen; C054234:chordin; D045726:Metalloproteases; D010447:Peptide Hydrolases; D055394:Tolloid-Like Metalloproteinases; D002918:Chymotrypsin; D008666:Metalloendopeptidases; C484478:TLL1 protein, human; C525705:BMP1 protein, human; D055395:Bone Morphogenetic Protein 1; D002118:Calcium