Sanderson I R, Xu Z, Chu S W, Xie Q Y, Levine L J, Walker W A
Developmental Gastroenterology Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, USA.
Gut. 1996 Jun;38(6):853-8. doi: 10.1136/gut.38.6.853.
The stimulatory guanosine triphosphate (GTP) binding protein alpha subunit (Gs alpha) of adenylate cyclase is the target protein for cholera toxin.
AIMS/METHODS: The expression of this signal transducer was analysed in the small intestine of developing rats by RNA transfer (northern blot) analysis by immunoblotting, and by ADP-ribosylation of membrane proteins.
Intestinal Gs alpha mRNA (about 1.9 kb) was increased in the neonate compared with the adult rat. Two isoforms of Gs alpha proteins, a 45,000 and a 52,000 form, were expressed in the small intestinal epithelial cell and both were ADP-ribosylated by cholera toxin. A significant increase in the larger isoform (52,000) and in its ribosylation was noted in the 2 week old suckling compared with post-weaned older animals. The protein content or ribosylation of the smaller form (45,000) did not significantly change with age.
These data show that a developmental decline of intestinal Gs alpha expression seems to be, in part, regulated at the mRNA level. An increased Gs alpha expression in the immature intestine may help to explain a previously reported, dose dependent increased adenylate cyclase response and an increase in fluid secretion to cholera toxin in neonates compared with adults.
腺苷酸环化酶的刺激性鸟苷三磷酸(GTP)结合蛋白α亚基(Gsα)是霍乱毒素的靶蛋白。
目的/方法:通过RNA转移(Northern印迹)分析、免疫印迹以及膜蛋白的ADP核糖基化,对发育中大鼠小肠中这种信号转导蛋白的表达进行分析。
与成年大鼠相比,新生大鼠小肠中的Gsα mRNA(约1.9 kb)有所增加。在小肠上皮细胞中表达了两种Gsα蛋白异构体,一种45000形式和一种52000形式,两者均被霍乱毒素进行ADP核糖基化。与断奶后的年长动物相比,在2周龄的乳鼠中较大的异构体(52000)及其核糖基化显著增加。较小形式(45000)的蛋白质含量或核糖基化并未随年龄显著变化。
这些数据表明,肠道Gsα表达的发育性下降似乎部分在mRNA水平受到调节。未成熟肠道中Gsα表达的增加可能有助于解释先前报道的,与成年人相比,新生儿中腺苷酸环化酶反应呈剂量依赖性增加以及对霍乱毒素的液体分泌增加的现象。