Quaroni A, Kirsch K, Herscovics A, Isselbacher K J
Biochem J. 1980 Oct 15;192(1):133-44. doi: 10.1042/bj1920133.
The biosynthesis of membrane proteins and glycoproteins has been studied in rat intestinal crypt and villus cells by measuring the incorporation of L-[5,6-3H] fucose, D-[2-3H] mannose and L-[3,4,5-3H] leucine, given intraperitoneally, into Golgi, lateral-basal and luminal membranes. Incorporation of leucine and mannose was approximately equal in crypt and villus cells, whereas fucose incorporation was markedly higher (3-4 times) in the differentiated villus cells. As previously reported [Quaroni, Kirsch & Weiser (1979) Biochem J. 182. 203-212] most of the fucosylated glyco-proteins synthesized in the villus cells and initially present in the Golgi and lateral-basal membranes were found re-distributed, within 3-4h of label administration, in the luminal membrane. A similar process appeared to occur in the crypt cells, where, however, only few fucose-labelled glycoproteins were identified. In contrast, most of the leucine-labelled and many mannose-labelled membrane components found in the lateral-basal membrane of both crypt and villus cells did not seen to undergo a similar re-distribution process. The fucosylated glycoproteins of the intestinal epithelial cells represent, therefore, a special class of membrane components, most of which appear with differentiation, that are selectively localized in the luminal portion of the plasmalemma. In contrast with the marked differences in protein and glycoprotein patterns between the luminal membrane of villus and crypt cells, only minor differences were found between their lateral-basal membrane components: their protein patterns on sodium dodecyl sulphate/polyacrylamide slab gels, and the patterns of fucose-, mannose- and leucine-labelled components (analysed 3-4h after label administration) were very similar. Although the minor differences detected may be of importance, it appears that most of the surface-membrane changes accompanying cell differentiation in the intestinal epithelial cells are localized in the luminal portion of their surface membrane.
通过测量腹腔注射的L-[5,6-³H]岩藻糖、D-[2-³H]甘露糖和L-[3,4,5-³H]亮氨酸掺入大鼠小肠隐窝和绒毛细胞的高尔基体膜、侧基膜和腔面膜中的情况,对膜蛋白和糖蛋白的生物合成进行了研究。亮氨酸和甘露糖在隐窝细胞和绒毛细胞中的掺入量大致相等,而岩藻糖在分化的绒毛细胞中的掺入量明显更高(3至4倍)。如先前报道[Quaroni、Kirsch和Weiser(1979年)《生物化学杂志》182卷,203 - 212页],在绒毛细胞中合成并最初存在于高尔基体膜和侧基膜中的大多数岩藻糖化糖蛋白,在给予标记物后3至4小时内,被发现重新分布到了腔面膜中。类似的过程似乎也发生在隐窝细胞中,然而,在隐窝细胞中仅鉴定出少量岩藻糖标记的糖蛋白。相比之下,在隐窝和绒毛细胞的侧基膜中发现的大多数亮氨酸标记和许多甘露糖标记的膜成分似乎并未经历类似的重新分布过程。因此,肠上皮细胞的岩藻糖化糖蛋白代表了一类特殊的膜成分,其中大多数随着分化而出现,它们选择性地定位于质膜的腔面部分。与绒毛和隐窝细胞腔面膜之间蛋白质和糖蛋白模式的显著差异相反,在它们的侧基膜成分之间仅发现了微小差异:它们在十二烷基硫酸钠/聚丙烯酰胺平板凝胶上的蛋白质模式,以及岩藻糖、甘露糖和亮氨酸标记成分的模式(在给予标记物后3至4小时分析)非常相似。尽管检测到的微小差异可能很重要,但似乎肠上皮细胞中伴随细胞分化的大多数表面膜变化都局限于其表面膜的腔面部分。