Felipe A, Remesar X, Pastor-Anglada M
Departament de Bioquímica i Fisiologia, Universitat de Barcelona, Spain.
Am J Physiol. 1995 Mar;268(3 Pt 2):R598-604. doi: 10.1152/ajpregu.1995.268.3.R598.
Changes in L-alanine transport in plasma membrane vesicles from livers of control and 24- and 48-h starved adult rats and the sensitivity of alanine uptake to sulfhydryl group reagents [N-ethylmaleimide (NEM) and p-chloromercuribenzenesulfonate (p-CMBS)] were studied. The portal concentration of certain amino acids was measured, and the relationship between L-alanine transport kinetic parameters and amino acid levels was analyzed. Starvation only induced a decrease in portal concentration of these amino acids that are mainly carried by Na(+)-dependent systems (85 and 61% for 24- and 48-h starved rats, respectively). Portal alanine concentration was lower in 24-h starved animals than in control rats (370 vs. 587 microM) and further decreased after 48 h of fasting (228 microM). Starvation induced an increase in maximum velocity (Vmax) values of Na(+)-dependent L-alanine transport (7.19, 8.97, and 12.38 pmol.U 5'-nucleotidase-1.10 s-1 for control and 24- and 48-h starved rats, respectively) with slight, but not significant, changes in the apparent Michaelis constant (Km) values (3.35, 2.63, and 2.20 mM for control and 24- and 48-h starved rats, respectively). Portal alanine showed a directly close correlation with Km values and inverse with Vmax values. The mean affinity constant values for the effects of NEM and p-CMBS on Na(+)-dependent L-alanine transport were lower in 48- (2.57 and 0.13 mM, respectively) and 24-h starved rats (3.59 and 0.32 mM, respectively) than in control rats (8.56 and 0.59 mM, respectively) and showed a directly strong correlation with kinetic characteristics of L-alanine transport and portal alanine concentration.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了对照成年大鼠以及饥饿24小时和48小时的成年大鼠肝脏质膜囊泡中L-丙氨酸转运的变化,以及丙氨酸摄取对巯基试剂[N-乙基马来酰亚胺(NEM)和对氯汞苯磺酸盐(p-CMBS)]的敏感性。测量了某些氨基酸的门静脉浓度,并分析了L-丙氨酸转运动力学参数与氨基酸水平之间的关系。饥饿仅导致主要由Na(+)依赖性系统转运的这些氨基酸的门静脉浓度降低(饥饿24小时和48小时的大鼠分别降低85%和61%)。饥饿24小时的动物门静脉丙氨酸浓度低于对照大鼠(370 μM对587 μM),禁食48小时后进一步降低(228 μM)。饥饿导致Na(+)依赖性L-丙氨酸转运的最大速度(Vmax)值增加(对照大鼠、饥饿24小时和48小时的大鼠分别为7.19、8.97和12.38 pmol·U 5'-核苷酸酶-1·10 s-1),表观米氏常数(Km)值有轻微但不显著的变化(对照大鼠、饥饿24小时和48小时的大鼠分别为3.35、2.63和2.20 mM)。门静脉丙氨酸与Km值呈直接密切相关,与Vmax值呈负相关。NEM和p-CMBS对Na(+)依赖性L-丙氨酸转运作用的平均亲和常数在饥饿48小时(分别为2.57和0.13 mM)和24小时的大鼠(分别为3.59和0.32 mM)中低于对照大鼠(分别为8.56和0.59 mM),并且与L-丙氨酸转运的动力学特征和门静脉丙氨酸浓度呈直接强相关。(摘要截断于250字)