Human nidogen-2 was cloned and sequenced (1375 residues) and found to share 46% sequence identity and a similar domain arrangement with the previously characterized basement membrane protein nidogen-1. Recombinant nidogen-2 was purified as a 200 kDa protein from transfected mammalian cell medium, showed a high level of N and O-glycosylation, and could be clearly distinguished from nidogen-1 (150 kDa) by specific antibodies. Electron microscopy demonstrated that the two isoforms have a similar shape, consisting of three globular domains connected by two threads, but differ somewhat in length. Northern blots and immunological assays demonstrated co-expression of the nidogens in various tissues and cultured cells. Immunofluoresence revealed colocalization in vessel walls and other basement membrane zones but some differences in heart and skeletal muscle. Nidogen-2 interacted with collagens I and IV, and perlecan at a comparable level to nidogen-1 but failed to bind to fibulins. Nidogen-2 bound to laminin-1, but only moderately to the epitope on the laminin gamma1 chain, which promotes high-affinity binding of nidogen-1. Both nidogens were cell-adhesive for a restricted number of cell lines, with nidogen-2 having a higher activity. Together, these data suggest that nidogen-2 can compensate for some but not all functional activities ascribed to nidogen-1.
Copyright 1998 Academic Press.