Bonen A, Luiken J J, Liu S, Dyck D J, Kiens B, Kristiansen S, Turcotte L P, Van Der Vusse G J, Glatz J F
Department of Kinesiology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1.
Am J Physiol. 1998 Sep;275(3):E471-8. doi: 10.1152/ajpendo.1998.275.3.E471.
We performed studies 1) to investigate the kinetics of palmitate transport into giant sarcolemmal vesicles, 2) to determine whether the transport capacity is greater in red muscles than in white muscles, and 3) to determine whether putative long-chain fatty acid (LCFA) transporters are more abundant in red than in white muscles. For these studies we used giant sarcolemmal vesicles, which contained cytoplasmic fatty acid binding protein (FABPc), an intravesicular fatty acid sink. Intravesicular FABPc concentrations were sufficiently high so as not to limit the uptake of palmitate under conditions of maximal palmitate uptake (i.e., 4.5-fold excess in white and 31.3-fold excess in red muscle vesicles). All of the palmitate taken up was recovered as unesterified palmitate. Palmitate uptake was reduced by phloretin (-50%), sulfo-N-succinimidyl oleate (-43%), anti-plasma membrane-bound FABP (FABPpm, -30%), trypsin (-45%), and when incubation temperature was lowered to 0 degrees C (-70%). Palmitate uptake was also reduced by excess oleate (-65%), but not by excess octanoate or by glucose. Kinetic studies showed that maximal transport was 1.8-fold greater in red vesicles than in white vesicles. The Michaelis-Menten constant in both types of vesicles was approximately 6 nM. Fatty acid transport protein mRNA and fatty acid translocase (FAT) mRNA were about fivefold greater in red muscles than in white muscles. FAT/CD36 and FABPpm proteins in red vesicles or in homogenates were greater than in white vesicles or homogenates (P < 0.05). These studies provide the first evidence of a protein-mediated LCFA transport system in skeletal muscle. In this tissue, palmitate transport rates are greater in red than in white muscles because more LCFA transporters are available.
1)探究棕榈酸转运至巨大肌膜囊泡的动力学;2)确定红色肌肉中的转运能力是否高于白色肌肉;3)确定假定的长链脂肪酸(LCFA)转运蛋白在红色肌肉中是否比在白色肌肉中更丰富。在这些研究中,我们使用了含有细胞质脂肪酸结合蛋白(FABPc)的巨大肌膜囊泡,FABPc是一种囊泡内脂肪酸汇。囊泡内FABPc浓度足够高,以至于在最大棕榈酸摄取条件下(即白色肌肉囊泡中过量4.5倍,红色肌肉囊泡中过量31.3倍)不会限制棕榈酸的摄取。摄取的所有棕榈酸均以未酯化的棕榈酸形式回收。根皮素(-50%)、磺基-N-琥珀酰亚胺油酸酯(-43%)、抗质膜结合FABP(FABPpm,-30%)、胰蛋白酶(-45%)以及将孵育温度降至0摄氏度(-70%)时,棕榈酸摄取均减少。过量油酸(-65%)也会降低棕榈酸摄取,但过量辛酸或葡萄糖则不会。动力学研究表明,红色囊泡中的最大转运量比白色囊泡大1.8倍。两种类型囊泡中的米氏常数约为6 nM。红色肌肉中脂肪酸转运蛋白mRNA和脂肪酸转位酶(FAT)mRNA比白色肌肉中大约高五倍。红色囊泡或匀浆中的FAT/CD36和FABPpm蛋白高于白色囊泡或匀浆(P < 0.05)。这些研究首次证明了骨骼肌中存在蛋白质介导的LCFA转运系统。在该组织中,红色肌肉中的棕榈酸转运速率高于白色肌肉,因为有更多的LCFA转运蛋白。