ISOLATION OF HUMAN SKELETAL MUSCLE MYOSIN HEAVY CHAIN & ACTIN

我们开发了一种分离肌苷重链的简单方法 (MHC)和肌动蛋白从小(60-80毫克)人骨骼肌样本中提取 用于测定它们的分数合成速率，利用 聚丙烯酰胺凝胶电泳(PAGE)。MHC和MHC的含量 肌动蛋白可用于[13C]亮氨酸的定量 气相色谱-燃烧-同位素比质法测定丰度 光谱(GC/C/IRMS)。混合肌肉的分数合成率 对6名健康青年进行了蛋白质、MHC和肌动蛋白的测定 受试者在接受14小时静脉输液后27(3年) (Prime=7.58(mol.kg-1，恒定输液量=7.58(mol.kg-1.hr-1)) [1-13C]亮氨酸。总体而言，MHC的合成率降低了20% 肌动蛋白合成率提高61%(P=0.060)。 (N.S.)而不是基质金属蛋白酶的合成率。这些蛋白质的分离 为利用GC/C/IRMS进行同位素丰度分析提供了有价值的参考 关于这些S特异缩合物的合成速率的信息 蛋白质，而不是由 以往对基质金属蛋白酶合成率的研究。

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.