Inter- and intramolecular disulfide bonding among lymphocyte plasma membrane proteins and glycoproteins.
The disulfide bonding characteristics of the pig lymph node plasma membrane (PM) proteins and glycoproteins have been examined by 1- and 2-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Reaction of the purified PM vesicles with N-ethyl maleimide (NEM) prior to detergent solubilization was found to markedly reduce the extent of intermolecular disulfide bonding subsequently observed. Thus the blocking of free sulfhydryl groups with NEM prevented the detergent-induced disulfide bonding of numerous components, including PM-bound actin. The extent of intermolecular disulfide bonding among the NEM-pretreated PM glycoproteins purified by lentil lectin affinity chromatography was found to be relatively limited, with only 3% of the total glycoprotein present as intermolecular disulfide-bonded complexes. In contrast, the degree of intramolecular disulfide bonding revealed by a modified 1-dimensional SDS-PAGE technique was quite striking. Among those polypeptides demonstrating a clearly altered mobility upon reduction was the heavy chain of class I and beta-chain of class II major histocompatibility complex (MHC) antigens. The class II alpha-chain, however, was much less affected. These changes have been compared with those observed for proteins containing intramolecular disulfide-bonded domains of known size and number, and considered in the light of recent information on the structure of MHC antigens.
D000818:Animals; D004220:Disulfides / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D006023:Glycoproteins / Q000032:analysis*; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000032:analysis* / Q000187:drug effects / Q000276:immunology; D046911:Macromolecular Substances; D008285:Major Histocompatibility Complex; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D011487:Protein Conformation; D013552:Swine
D004220:Disulfides; D006023:Glycoproteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D005033:Ethylmaleimide