Effect of dithiothreitol on the 1 alpha, 25-dihydroxyvitamin D3 chick intestinal receptor analyzed by polyacrylamide gel electrophoresis.
This is the first report of the use of non-denaturing polyacrylamide gel electrophoresis (PAGE) to measure the apparent molecular weight of the chick intestinal 1 alpha, 25-dihydroxyvitamin D3 (1, 25-(OH)2 - D3) receptor and to study the effect of dithiothreitol on it. When prepared in the absence of this factor, chick intestinal cytosol contained one major specific 1,25 - (OH)2 - D3 binding peak. Its apparent molecular weight was 95,200 +/- 1,900 (SD) daltons. Preparation of the cytosol in the presence of 5 mM dithiothreitol resulted in the appearance, besides the 95,000 daltons peak, of an additional 1,25 - (OH)2 - D3 binding peak, the molecular weight of which was 73,600 +/- 3,300 (SD). This effect of dithiothreitol could be suppressed by the simultaneous addition of 10 mM N alpha-p-tosyl-L-arginine methyl ester (TAME), a protease inhibitor.
D000818:Animals; D002645:Chickens; D003600:Cytosol / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D007413:Intestinal Mucosa / Q000378:metabolism*; D008297:Male; D008970:Molecular Weight; D018167:Receptors, Calcitriol; D011987:Receptors, Steroid / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*
D018167:Receptors, Calcitriol; D011987:Receptors, Steroid; D004229:Dithiothreitol