Hoffman L R, Chang E B
Department of Medicine, University of Chicago, Illinois 60637.
J Biol Chem. 1991 Nov 15;266(32):21815-20.
Sucrase-isomaltase (SI) expression along the longitudinal and vertical axis of the small intestine was studied by sequentially isolating enterocytes from villus to crypt of rat proximal jejunum and distal ileum. Gradients of sucrase activity were observed with greatest activity occurring in jejunal and villus regions. Along the villus-to-crypt axis, gradients of SI mRNA abundance corresponded with activity. However, along the longitudinal axis no differences in SI mRNA levels were observed, thus not accounting for the observed 3-5-fold difference in SI activities between jejunum and ileum. Comparison of SI immunoprecipitates from jejunal and ileal mucosal scrapings showed significant differences in gel mobilities of the more mature forms, which did not appear to affect SI functional activities. When relative rates of de novo SI protein synthesis were compared, [35S]methionine incorporation into all SI forms was observed to be 3-5-fold greater in jejunum than in ileum at all time points. Because these results suggested differences in regional translational regulation, subcellular distribution of SI mRNA in jejunal and ileal epithelial cells was compared. A greater proportion of jejunal SI mRNA was found to be associated with membrane-bound polyribosomes. We conclude 1) sucrase expression along the villus-to-crypt axis correlates with SI mRNA abundance, 2) post-translational processing of SI differ in ileum and jejunum, but appear not to determine SI expression, and 3) differences in translational processing in distal ileum and proximal jejunum may determine sucrase activity along the longitudinal axis of rat small intestine.
通过依次从大鼠近端空肠和远端回肠的绒毛到隐窝分离肠上皮细胞,研究了蔗糖酶 - 异麦芽糖酶(SI)在小肠纵轴和垂直轴上的表达。观察到蔗糖酶活性梯度,在空肠和绒毛区域活性最高。沿着绒毛到隐窝轴,SI mRNA丰度梯度与活性相对应。然而,沿着纵轴未观察到SI mRNA水平的差异,因此无法解释空肠和回肠之间观察到的SI活性3至5倍的差异。对空肠和回肠黏膜刮片的SI免疫沉淀产物进行比较,发现更成熟形式的凝胶迁移率存在显著差异,这似乎并未影响SI的功能活性。比较从头合成SI蛋白的相对速率时,在所有时间点均观察到,[35S]甲硫氨酸掺入空肠中所有SI形式的量比回肠高3至5倍。由于这些结果提示了区域翻译调控的差异,因此比较了空肠和回肠上皮细胞中SI mRNA的亚细胞分布。发现空肠中更大比例的SI mRNA与膜结合多核糖体相关。我们得出结论:1)沿着绒毛到隐窝轴的蔗糖酶表达与SI mRNA丰度相关;2)SI在回肠和空肠中的翻译后加工不同,但似乎不决定SI的表达;3)远端回肠和近端空肠中翻译加工的差异可能决定大鼠小肠纵轴上的蔗糖酶活性。