Han X, Budreau A M, Chesney R W
Department of Pediatrics, University of Tennessee, Memphis 38103, USA.
Pediatr Res. 1997 May;41(5):624-31. doi: 10.1203/00006450-199705000-00004.
Renal brush border taurine transport adapts to changes in the dietary intake of sulfur amino acids with increased rates after dietary restriction and reduced transport after dietary surplus. The Xenopus laevis oocyte expression system was used to define the renal adaptive response to dietary manipulation. Injection of poly(A)+ RNA isolated from rat kidney cortex resulted in a time- and dose-dependent increase in NaCl-taurine cotransport in oocytes. The Km of the expressed taurine transporter was 22.5 microM. In oocytes, injection of 40 ng of poly(A)+ RNA from kidneys of low taurine diet (LTD)-fed rats elicited 2-fold the taurine uptake of normal taurine diet (NTD)-fed rats and >3-fold the uptake of high taurine diet (HTD)-fed rats. Northern blots of rat kidneys using a riboprobe derived from an rB16a (rat brain taurine transporter) subclone revealed 6.2- and 2.4-kb transcripts, the abundance of which were increased or decreased in LTD- or HTD-fed rats, respectively, as compared with NTD-fed rats. A approximately 70-kD protein was detected by Western blot using an antibody derived from a synthetic peptide corresponding to a conserved intracellular segment of rB16a. The abundance of the approximately 70-kD protein was increased or decreased in LTD- or HTD-fed rats, respectively, as compared with NTD-fed rats. In conclusion, expression of the rat renal taurine transporter is regulated by dietary taurine at the level of mRNA accumulation and protein synthesis.
肾刷状缘牛磺酸转运可适应含硫氨基酸饮食摄入量的变化,饮食限制后转运速率增加,饮食过剩后转运减少。非洲爪蟾卵母细胞表达系统用于确定肾脏对饮食调控的适应性反应。注射从大鼠肾皮质分离的多聚腺苷酸加尾RNA(poly(A)+ RNA)会使卵母细胞中氯化钠 - 牛磺酸共转运呈时间和剂量依赖性增加。所表达的牛磺酸转运体的米氏常数(Km)为22.5微摩尔。在卵母细胞中,注射来自低牛磺酸饮食(LTD)喂养大鼠肾脏的40纳克poly(A)+ RNA,其牛磺酸摄取量是正常牛磺酸饮食(NTD)喂养大鼠的2倍,是高牛磺酸饮食(HTD)喂养大鼠摄取量的3倍以上。用源自rB16a(大鼠脑牛磺酸转运体)亚克隆的核糖探针进行大鼠肾脏的Northern印迹分析,显示有6.2千碱基和2.4千碱基的转录本,与NTD喂养大鼠相比,LTD或HTD喂养大鼠中这些转录本的丰度分别增加或减少。使用源自与rB16a保守细胞内区段对应的合成肽的抗体进行Western印迹分析,检测到一种约70千道尔顿的蛋白质。与NTD喂养大鼠相比,LTD或HTD喂养大鼠中这种约70千道尔顿蛋白质的丰度分别增加或减少。总之,大鼠肾脏牛磺酸转运体的表达在mRNA积累和蛋白质合成水平受饮食中牛磺酸的调节。