Role of Acyl-CoA Synthetase-5 in Fatty Acid Partitioning

描述（由申请方提供）：营养相关疾病如肥胖症和糖尿病表现出影响β-氧化和三酰甘油储存的酰基辅酶A分配缺陷。酰基辅酶A合成酶（ACS）催化脂肪酸合成的第一步。本研究的目的是确定ACS 5异构体在脂肪酸分配途径能量产生和甘油脂质合成之间的作用。ACS 5和ACS 5反义mRNA将通过重组腺病毒技术在原代肝细胞培养物中过表达。将测量14-C油酸酯掺入β氧化和脂质合成产物中，以确定过量或降低的酶水平对途径的影响。此外，ACS 5将在从缺乏功能线粒体甘油-3-磷酸酰基转移酶（GPAT）的小鼠中分离的原代肝细胞中表达，GPAT催化甘油脂质合成的初始步骤。这些实验将确定过量或降低的ACS 5酶水平是否会在存在和不存在GPAT的情况下改变β-氧化和脂质合成的速率，降低激素刺激的代谢效应，或在β-氧化被抑制时增加脂质合成。我们假设，ACS 5参与脂肪酸分配提供酰基辅酶A主要用于能量生产的β-氧化。

DESCRIPTION (provided by applicant): Nutrition-related diseases such as obesity and diabetes mellitus exhibit defective acyl-CoA partitioning which affects beta-oxidation and triacylglycerol stores. Acyl CoA synthetase (ACS) catalyzes the first step in fatty acid The purpose of this study is to identify the role of the ACS5 isoform in fatty acid partitioning between pathway energy production and glycerolipid synthesis. ACS5 and ACS5 antisense mRNA will be overexpressed in primary hepatocyte cultures via recombinant adenoviral technology. Incorporation of 14-C oleate into products of beta oxidation and lipid synthesis will be measured to determine the effects of excess or reduced enzyme levels on pathways. Additionally, ACS5 will be expressed in primary hepatocytes isolated from mice lacking function mitochondrial glycerol-3-phosphate acyltransferase (GPAT), which catalyzes the initial step in glycerolipid synthesis. These experiments will determine if excess or reduced enzyme levels of ACS5 alter rates of beta-oxidation and lipid synthesis in the presence and absence of GPAT, reduce the metabolic effects of hormonal stimuli, or increase lipid synthesis when beta-oxidation is inhibited. We hypothesize that ACS5 participates in fatty acid partitioning by providing acyl-CoAs primarily for energy production by beta-oxidation.