Characterization of high affinity progesterone-binding membrane proteins by anti-peptide antiserum.
A chemically synthesized 15-mer oligopeptide derived from the N terminus of high affinity progesterone-binding membrane site(s) from porcine liver was used to generate site-specific antibodies. Western blotting experiments confirmed the specificity of the anti-peptide serum obtained. In further investigations this antiserum was used for the identification of the native progesterone-binding membrane protein complex that represents an oligomer with an apparent molecular mass of about 200 kDa. In temperature-induced Triton X-114 phase separation experiments combined with Western-blotting, the progesterone-binding site was identified as an hydrophobic (integral) membrane protein. In addition, in Western blotting analyses the antiserum reacted with the progesterone-binding or related proteins in membrane fractions from a wide array of different tissues in various species.
Meyer
C
C
Division of Clinical Pharmacology, Klinikum Innenstadt, University of Munich, Germany.
Schmid
R
R
Schmieding
K
K
Falkenstein
E
E
Wehling
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D015153:Blotting, Western; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007425:Intracellular Membranes / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D008862:Microsomes, Liver / Q000648:ultrastructure; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D011374:Progesterone / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013552:Swine
D007106:Immune Sera; D008565:Membrane Proteins; D010446:Peptide Fragments; D011980:Receptors, Progesterone; D011374:Progesterone