Adrenocortical pregnenolone-binding protein: identification and antibody development.
Pregnenolone-binding activity isolated from the cytosol of the guinea pig adrenal cortex appears to correspond to a Mr 34,000 protein when examined by SDS-polyacrylamide gel electrophoresis during different stages of purification. To verify this finding the Mr 34,000 protein band was eluted from the SDS gel and used to generate a polyclonal antibody. Immobilized anti 34,000 IgG on protein A-Sepharose was found to extract pregnenolone-binding activity from solution in contrast to pre-immune IgG and an antibody raised against a Mr 30,000 protein isolated simultaneously. In addition, protein eluted from the protein A-anti 34,000 IgG complex exhibited the expected molecular weight of 34,000 when examined on an SDS gel. These results, thus, confirm that the pregnenolone-binding protein is indeed a protein of Mr 34,000.
Lee
Y J
YJ
Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, Bethesda, Maryland 20892.
Strott
C A
CA
D000302:Adrenal Cortex / Q000032:analysis*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000941:Antigens / Q000276:immunology; D002352:Carrier Proteins / Q000276:immunology / Q000302:isolation & purification*; D002845:Chromatography; D003600:Cytosol / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D007118:Immunoassay; D007074:Immunoglobulin G; D008297:Male; D008970:Molecular Weight; D011284:Pregnenolone / Q000378:metabolism
D000906:Antibodies; D000941:Antigens; D002352:Carrier Proteins; D007074:Immunoglobulin G; D011284:Pregnenolone