Tessier S, Chapdelaine A, Chevalier S
Maisonneuve-Rosemont Research Center, University of Montreal, Quebec, Canada.
Mol Cell Endocrinol. 1989 Jun;64(1):87-94. doi: 10.1016/0303-7207(89)90068-3.
To evaluate the possible role of intracellular phosphatases in the local regulation of prostatic functions, the effect of sodium orthovanadate (VO4), an inhibitor of phosphotyrosyl protein phosphatases, was studied on both protein phosphorylation and acid phosphatase activity. Secretory and non-secretory epithelial cells were isolated from normal and metaplastic prostates and incubated with [32P]phosphate in the presence and in the absence of VO4; the phosphoproteins were separated by electrophoresis and the gels were either directly submitted to autoradiography or after an alkali treatment to reveal those proteins enriched in phosphotyrosine. Prior to alkali treatment, several phosphoproteins were evidenced and in less than half of the cell preparations a slight increase in labeling intensity under vanadate (less than 75%) was observed in two phosphoproteins, p57 and p44. After alkali treatment: (1) the effect of VO4 on p57 remained in the order of 44-45% and it was restricted to less than half of non-secretory cell preparations; (2) its effect on p44 was intensified (134-207%) and observed in all cell types and in more than 80% of all preparations; and (3) in half of non-secretory cell preparations from metaplastic glands, an effect of VO4 on p35 (127%) became evident. In all instances, with normal and/or metaplastic prostates, protein phosphorylation activity, either total or alkali-resistant and in the presence or in the absence of VO4, was always higher in non-secretory epithelial cells as compared to secretory cells.(ABSTRACT TRUNCATED AT 250 WORDS)
为评估细胞内磷酸酶在前列腺功能局部调节中的可能作用,研究了磷酸酪氨酸蛋白磷酸酶抑制剂原钒酸钠(VO4)对蛋白质磷酸化和酸性磷酸酶活性的影响。从正常和化生前列腺中分离出分泌性和非分泌性上皮细胞,在有和没有VO4的情况下用[32P]磷酸盐孵育;通过电泳分离磷蛋白,凝胶要么直接进行放射自显影,要么经过碱处理以揭示富含磷酸酪氨酸的蛋白质。在碱处理之前,已证实有几种磷蛋白,并且在不到一半的细胞制剂中,观察到在钒酸盐作用下两种磷蛋白p57和p44的标记强度略有增加(小于75%)。碱处理后:(1)VO4对p57的作用保持在44 - 45%的水平,且仅限于不到一半的非分泌性细胞制剂;(2)其对p44的作用增强(134 - 207%),在所有细胞类型以及超过80%的所有制剂中都能观察到;(3)在化生腺体的一半非分泌性细胞制剂中,VO4对p35的作用(127%)变得明显。在所有情况下,对于正常和/或化生前列腺,无论有无VO4,非分泌性上皮细胞中的蛋白质磷酸化活性,无论是总活性还是耐碱性活性,总是高于分泌性细胞。(摘要截短于250字)