INTERACTIONS OF THE AMINO-TERMINAL NONCOLLAGENOUS (NC1) DOMAIN OF TYPE-VII COLLAGEN WITH EXTRACELLULAR-MATRIX COMPONENTS - A POTENTIAL ROLEIN EPIDERMAL-DERMAL ADHERENCE IN HUMAN SKIN

Type VII collagen, the major component of anchoring fibrils, consists of a central collagenous triple-helical domain flanked by two noncolla genous domains, NC1 and NC2. The NC1 domain contains multiple submodul es with homology to known adhesive molecules including fibronectin typ e III-like repeats and the A domain of von Willebrand factor. In this study, we produced the entire NC1 domain of human type VII collagen in the stably transfected human kidney 293 cell clones and purified larg e quantities of the recombinant NC1 protein from serum-free culture me dia. The recombinant NC1 formed interchain disulfide-bonded dimers and trimers and was N-linked glycosylated. Tunicamycin inhibited the cell ular secretion of NC1, suggesting that N-linked glycosylation may play a role in NC1 secretion, The recombinant NC1 was indistinguishable fr om the authentic NCI obtained from human amnions or WISH cells with re spect to N-linked sugar content, electrophoretic mobility, rotary shad ow imaging, and binding affinity to type IV collagen. Purified recombi nant NC1, like authentic NC1, also bound specifically to fibronectin, collagen type I, and a laminin 5/6 complex. Both monomeric and trimeri c forms of NC1 exhibited equal affinity for these extracellular matrix components, suggesting that the individual arms of NC1 can function i ndependently. The multiple interactions of NC1 with other extracellula r matrix components may support epidermal-dermal adhesion.