Physical and immunochemical studies of the globular domain of type IV collagen. Cryptic properties of the Goodpasture antigen.
The globular domain of type IV collagen from bovine glomerular basement membrane was isolated under nondenaturing conditions. It was shown to exist in a hexameric form comprising monomeric and dimeric subunits, with the Goodpasture antigen residing in monomer M2 and dimer D2 as previously described (Butkowski, R. J., Wieslander, J., Wisdom, B. J., Barr, J. F., Noelken, M. E., and Hudson, B. G. (1985) J. Biol. Chem. 260, 3739-3747). The epitope, however, is sequestered inside the hexamer, but becomes exposed and binds with the Goodpasture antibody upon dissociation of the hexamer into its subunits after treatment with concentrated guanidine HC1 or dilute acetic acid (pH less than 3.0). The process is completely reversible even from the denatured state. Circular dichroism studies show that the conformation of each subunit is unusually resistant to change in 6 M guanidine HC1 at 25 degrees C. This suggests that exposure of the epitope by dissociation requires minimal or no unfolding of subunits. The results provide additional evidence for localization of the Goodpasture antigen to the globular domain of type IV collagen. Moreover, these studies extend the conclusion (Weber, H., Engel, J., Wiedemann, H., Glanville, R., and Timpl, R. (1984) Eur. J. Biochem. 139, 401-410) about a tumor basement membrane, to an authentic physiological membrane, that the globular domain is a major cross-linking site in the type IV collagen matrix.
J
J
Wieslander
J
J
Langeveld
R
R
Butkowski
M
M
Jodlowski
M
M
Noelken
B G
BG
Hudson
D000818:Animals; D019867:Anti-Glomerular Basement Membrane Disease / Q000276:immunology*; D001485:Basement Membrane / Q000032:analysis; D002417:Cattle; D002942:Circular Dichroism; D003094:Collagen / Q000276:immunology*; D000939:Epitopes / Q000032:analysis*; D019791:Guanidine; D006146:Guanidines; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D007678:Kidney Glomerulus / Q000648:ultrastructure; D046911:Macromolecular Substances; D008854:Microscopy, Electron
D000939:Epitopes; D006146:Guanidines; D046911:Macromolecular Substances; D003094:Collagen; D019791:Guanidine