title: Large complex globular domains of type VII procollagen contribute to the structure of anchoring fibrils.
abstract: Type VII collagen, in the form of an antiparallel dimer, is a major protein component of anchoring fibrils. The ultrastructural appearance of these fibrils suggests that they may serve to anchor the basement membrane zone to the underlying connective tissue matrix. We report here the identification and initial characterization of Type VII procollagen, recovered from the media of epidermoid carcinoma cell cultures. Immunoblotting using monospecific antibodies to Type VII procollagen identifies a single, homogeneous band of at least Mr 320,000 following disulfide bond reduction. This chain contains 170 kDa of collagen triple helix and 150 kDa of non-helical domain at the carboxyl terminus. Pepsin digestion of this material yields Type VII collagen identical to that isolated from whole tissue and a series of quasi-stable peptides derived from the carboxyl-terminal region. Cell extracts contain procollagen chains identical in size to those secreted into the media. There is no evidence for processing of this material in cell culture. Partial purification by velocity sedimentation and transmission electron microscopic observation following rotary shadowing reveals both monomers (426 nm) and dimers (785 nm). Dimers are antiparallel and interact through 60-nm overlap, with amino-terminal globular domains present at the ends of the overlap. The multi-domain carboxyl-terminal region appears as three similar arms originating from a centralized globular region adjacent to the collagen helix. The carboxyl globular domain is present in whole tissue and may participate in the unique fibril form of this collagen. The amino-terminal globule may function in the antiparallel assembly of dimers.
mesh_terms: D000906:Antibodies; D005455:Fluorescent Antibody Technique; D006801:Humans; D007624:KB Cells / Q000378:metabolism; D046911:Macromolecular Substances; D003012:Microbial Collagenase / Q000378:metabolism; D008854:Microscopy, Electron; D008970:Molecular Weight; D010434:Pepsin A / Q000378:metabolism; D011347:Procollagen / Q000302:isolation & purification* / Q000378:metabolism; D011487:Protein Conformation
chemical_list: D000906:Antibodies; D046911:Macromolecular Substances; D011347:Procollagen; D010434:Pepsin A; D003012:Microbial Collagenase