Hormonal effects on the sulfhydryl groups associated with intestinal brush border membrane proteins

Abstract

Previous studies demonstrated that the administration of 1,25-dihydroxycholecalciferol (1,25(OH) 2D 3) to cholecalciferol-deficient chicks rapidly increases the reactivity and amount of the sulfhydryl (HS −) groups in intestinal brush border membranes (BBM). In the present study, the tissue and hormonal specificity of this effect was investigated. The HS − groups of intestinal and renal BBM were enhanced by vitamin D-3 and / or 1,25(OH) 2D 3, but no change was noted in isolated intestinal mitochondria and purified intestinal basolateral membranes, cardiac sarcolemma and erythrocyte membranes. Other steroid hormones including estradiol, testosterone, aldosterone, cortisol, dexamethasone and progesterone, yielded a response similar to 1,25(OH) 2D 3 on BBM HS − groups. Triiodothyronine and retinoic acid also resulted in an increase in intestinal BBM HS − groups. In a kinetic approach, using a specific sulfhydryl fluorescent probe ( N-7-dimethylamino-4-coumarin-3-yl-maleimide, DACM), the reactivity of the BBM HS − groups was increased by estrogen and testosterone, as was previously shown for 1,25(OH) 2D 3. Intestinal BBM proteins, labeled with DACM, were separated by gel electrophoresis. Fluorescence scans of the gel showed two heavily labeled bands, one of 110 kDa, putatively brush border myosin I, and one of 43 kDa, putatively actin. Labeling of the 110 kDa protein was increased by 1,25(OH) 2D 3 and estradiol. Further studies are required to elucidate the physiological meaning of these hormone-mediated increases in reactivity and amount of the BBM sulfhydryl groups, as well as the nature of the intermediate biochemical reactions involved in this response.