Wali R K, Bissonnette M, Skarosi S, Khare S, Sitrin M, Brasitus T A
Department of Medicine, University of Chicago, Chicago, Illinois, 60637, USA.
Biochem Biophys Res Commun. 1998 Sep 8;250(1):48-52. doi: 10.1006/bbrc.1998.9260.
Prior studies by our laboratory have shown that 1, 25-dihydroxyvitamin D3 activated PKC-alpha, but not PKC-delta, -epsilon, or -zeta, in normal rat colonocytes. In the present studies we demonstrate for the first time that this secosteroid also activated PKC-betaII, another DAG- and Ca2+-dependent PKC isoform recently shown to be present in these cells. Moreover, this activation of PKC-betaII by 1,25-dihydroxyvitamin D3 treatment of isolated colonocytes was shown to be lost in cells from vitamin D-deficient rats and, at least partially, restored by repleting these animals with this secosteroid for 7 days. Under basal conditions, the expression of PKC-alpha and -betaII in brush-border membranes was comparable to their respective expression in basolateral plasma membranes of rat colonocytes. In contrast, the expression of PKC-delta was significantly greater in brush-border membranes, whereas PKC-epsilon and -zeta were enriched in the basolateral plasma membranes. Furthermore, 1,25-dihydroxyvitamin D3 specifically induced the translocation of PKC-betaII, but not PKC-alpha, to the basolateral, but not brush-border plasma membranes of rat colonocytes, via a pp60(c-src)-dependent mechanism.
我们实验室之前的研究表明,在正常大鼠结肠细胞中,1,25 - 二羟基维生素D3可激活蛋白激酶Cα(PKC-α),但不能激活蛋白激酶Cδ(PKC-δ)、蛋白激酶Cε(PKC-ε)或蛋白激酶Cζ(PKC-ζ)。在本研究中,我们首次证明这种甾醇类激素还可激活PKC-βII,PKC-βII是另一种依赖二酰基甘油(DAG)和钙离子(Ca2+)的蛋白激酶C亚型,最近发现它也存在于这些细胞中。此外,用1,25 - 二羟基维生素D3处理分离的结肠细胞可激活PKC-βII,但在维生素D缺乏大鼠的细胞中这种激活作用消失,而通过给这些动物补充这种甾醇类激素7天,这种激活作用至少部分得以恢复。在基础条件下,PKC-α和PKC-βII在刷状缘膜中的表达与它们在大鼠结肠细胞基底外侧质膜中的各自表达相当。相反,PKC-δ在刷状缘膜中的表达明显更高,而PKC-ε和PKC-ζ则富集于基底外侧质膜中。此外,1,25 - 二羟基维生素D3通过一种依赖pp60(c-src)的机制,特异性地诱导PKC-βII而非PKC-α转位至大鼠结肠细胞的基底外侧质膜而非刷状缘质膜。