Evidence for a short form of alpha 1(IV) as a major polypeptide in bovine lens capsule.
The extracts from bovine lens capsule with acetic acid contained, after reduction, three major collagenous polypeptides with M(r) = 180k, 175k, and 160k, which were specifically immuno-stained with anti-type IV collagen polyclonal antibody. The biochemical properties of 180k and 160k polypeptides were akin and were distinct from that of 175k polypeptide [J. Biochem. (1993) 114,358-362]. In the present study, evidence that the 160k and 180k polypeptides from bovine lens capsule both originated from alpha 1(IV) was obtained on the basis of reactivity with a monoclonal antibody that recognizes alpha 1(IV) chain at the collagenous sequence contained in [KGEPGLPGRGFPGFP]. The epitope-bearing sequence was identified from the following three experiments. Pepsin-solubilized polypeptides from human placenta were purified by affinity chromatography on the antibody-coupled column and sequenced. The restriction map of the clones positively reactive with the monoclonal antibody from human placenta cDNA library was superimposed on that of human alpha 1(IV) cDNA at a specific region. Synthetic peptides corresponding to the sequence were assayed for inhibitory activity against the reaction between epitope-bearing pepsin fragments and the antibody. The 180k and 160k polypeptides showed similar intensities in protein staining as well as in immuno-staining with the monoclonal antibody. In contrast, the 175k polypeptide did not react with the monoclonal antibody, indicating that it is a genetically distinct type IV collagen chain, presumably alpha 2(IV) from its abundance. The 160k, a major type IV collagen polypeptide, is a short form of alpha 1(IV) present as a tissue form in bovine lens capsule.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D003094:Collagen / Q000032:analysis / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D015151:Immunoblotting; D007903:Lens Capsule, Crystalline / Q000737:chemistry*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D017421:Sequence Analysis
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010455:Peptides; D003094:Collagen