Stremmel W, Berk P D
Proc Natl Acad Sci U S A. 1986 May;83(10):3086-90. doi: 10.1073/pnas.83.10.3086.
When [14C]oleate bound to bovine serum albumin was incubated at 37 degrees C for 7 min with isolated rat hepatocytes in the absence of glucose, the cumulative oleate uptake curve had two components: a rapid, linear segment over the first 30 sec, followed by a slower, curvilinear component. At 173 microM [14C]oleate/albumin (1:1, mol/mol), the initial component had a slope (Vo) of 118 +/- 18 pmol per min per 5 X 10(4) hepatocytes (mean +/- SD). During this initial 30 sec, virtually no oleate was oxidized, and less than 11% was esterified. By 5 min, 79% was esterified; oxidation never exceeded 4%. Addition of 2 mM glucose significantly increased oleate esterification and thereby available oleate binding sites on cytosolic fatty acid binding protein but had no influence on Vo. Pretreatment with trypsin reduced Vo by 49 +/- 15%. These data indicate that the initial component of the oleate uptake curve reflects predominantly influx, whereas the subsequent component reflects a balance between influx, efflux, and intracellular metabolism. Influx is independent of intracellular binding, oxidation, and esterification and may reflect a membrane-associated carrier-mediated process.
当与牛血清白蛋白结合的[14C]油酸在无葡萄糖的情况下于37℃与分离的大鼠肝细胞一起孵育7分钟时,累积油酸摄取曲线有两个部分:在最初30秒内为快速线性部分,随后是较慢的曲线部分。在173微摩尔[14C]油酸/白蛋白(1:1,摩尔/摩尔)时,初始部分的斜率(Vo)为每5×10(4)个肝细胞每分钟118±18皮摩尔(平均值±标准差)。在最初的30秒内,几乎没有油酸被氧化,酯化的不到11%。到5分钟时,79%被酯化;氧化从未超过4%。添加2毫摩尔葡萄糖显著增加了油酸酯化,从而增加了胞质脂肪酸结合蛋白上可用的油酸结合位点,但对Vo没有影响。用胰蛋白酶预处理使Vo降低了49±15%。这些数据表明,油酸摄取曲线的初始部分主要反映流入,而随后的部分反映流入、流出和细胞内代谢之间的平衡。流入独立于细胞内结合、氧化和酯化,可能反映了一种与膜相关的载体介导过程。