Reductive cleavage of the disulfide bonds of the collagen IV noncollagenous domain in aqueous sodium dodecyl sulfate: absence of intermolecular nondisulfide cross-links.
The subunits of the collagen IV hexameric, noncollagenous NC1 domain obtained from bovine aorta, glomerular basement membrane, alveolar basement membrane and placental basement membrane are predominantly dimers. A large fraction of the dimers had been thought to be linked by nondisulfide bonds because they were resistant to cleavage by 5% (v/v) 2-mercaptoethanol, 2% (w/v) SDS, at 100 degrees C. However, if an unusually high concentration of 2-mercaptoethanol, e.g., 40% (v/v), is used, complete conversion of dimers into monomers is achieved, indicating the lack of intersubunit nondisulfide cross-links. Electrophoresis patterns indicate that some of the intermolecular disulfide bonds of the dimers are more resistant to reduction in aqueous SDS than are some of the intramolecular disulfide bonds.
Reddy
G K
GK
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66160-7421.
Hudson
B G
BG
Bailey
A J
AJ
Noelken
M E
ME
D000478:Alkylation; D000818:Animals; D001011:Aorta / Q000737:chemistry*; D001485:Basement Membrane / Q000737:chemistry; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000737:chemistry* / Q000302:isolation & purification; D003432:Cross-Linking Reagents; D004220:Disulfides / Q000737:chemistry*; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D006868:Hydrolysis; D007678:Kidney Glomerulus / Q000737:chemistry; D008239:Lysine; D046911:Macromolecular Substances; D008623:Mercaptoethanol; D009131:Muscle, Smooth, Vascular / Q000737:chemistry*; D010084:Oxidation-Reduction; D010920:Placenta / Q000737:chemistry; D011247:Pregnancy; D012967:Sodium Dodecyl Sulfate
D003432:Cross-Linking Reagents; D004220:Disulfides; D046911:Macromolecular Substances; D012967:Sodium Dodecyl Sulfate; D008623:Mercaptoethanol; D003094:Collagen; D008239:Lysine; D004229:Dithiothreitol