ISOLATION OF HUMAN SKELETAL MUSCLE MYOSIN HEAVY CHAIN & ACTIN

我们已经发展了一种简单的方法来分离mysoin重链 (MHC)和肌动蛋白从小（60-80毫克）的人骨骼肌样品 为了确定它们分数合成速率，利用 聚丙烯酰胺凝胶电泳（PAGE）。 MHC和 分离的肌动蛋白足以定量[13 C]亮氨酸 气相色谱-燃烧-同位素质量比丰度 质谱分析（GC/C/IRMS）。 混合肌合成率分数 对6名健康青年进行了基质金属蛋白酶（MMP）、MHC和肌动蛋白的测定 受试者（27例（3岁）接受14小时静脉输注后 （初始= 7.58（mol.kg-1），恒定输注= 7.58（mol.kg-1. hr-1） [1- 13 C]亮氨酸。 总的来说，MHC的合成率降低了20 （p=0.012）并且肌动蛋白的合成率高61%（p=0.060， N.S.）MMP的合成速率。 这些蛋白质的分离 GC/C/IRMS同位素丰度分析提供了有价值的 关于这些特异性收缩剂的合成速率的信息 蛋白质，而不是更一般的信息， MMP合成速率的先前研究。

We have developed a simple method to isolate mysoin heavy chain (MHC) and actin from small (60-80 mg) human skeletal muscle samples for determinatin of their fractional synthesis rates, utilizing polyacrylamide gel electrophoresis (PAGE). The amounts of MHC and actin isolated are adequate for the quantification of [13C]leucine abundance by gas chromatography-combustion-isotope ratio mass spectrometry (GC/C/IRMS). Fractional synthesis rates of mixed muscle protein (MMP), MHC, and actin were determined in six healthy young subjects (27 ( 3 yr) after they received a 14-hr intravenous infusion (prime = 7.58 (mol.kg-1, constant infusion = 7.58 (mol.kg-1.hr-1) of [1-13C]leucine. Overall, the synthesis rate of MHC was 20% lower (p=0.012) and the synthesis rate of actin was 61% higher (p=0.060, N.S.) than the MMP synthesis rate. The isolation of these proteins for isotope abundance analysis by GC/C/IRMS provides valuable information about the synthesis rates of these s pecific c ontractile proteins as opposed to the more general information provided by previous studies of MMP synthesis rates.