ISOLATION OF HUMAN SKELETAL MUSCLE MYOSIN HEAVY CHAIN & ACTIN

人体骨骼肌肌球蛋白重链的分离

• 批准号: 6665822
• 负责人: DL HASTEN
• 金额: $ 15.75万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6665822
• 关键词: aging; biological products; biomedical resource; female; human subject; human tissue; male; musculoskeletal system; physical fitness; proteins; spectrometry

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.

我们开发了一种分离肌球蛋白重链的简单方法 来自小量（60-80 毫克）人类骨骼肌样品的 (MHC) 和肌动蛋白 为了确定它们的分数合成率，利用 聚丙烯酰胺凝胶电泳（PAGE）。 MHC 的量和 分离的肌动蛋白足以定量[13C]亮氨酸 气相色谱-燃烧-同位素比质量丰度 光谱分析（GC/C/IRMS）。 混合肌肉的分数合成率 在六名健康年轻人中测定了蛋白质 (MMP)、MHC 和肌动蛋白 接受 14 小时静脉输注后的受试者（27 岁（3 岁）） （初始= 7.58（mol.kg-1，恒定输注= 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% (p=0.012) 肌动蛋白的合成率提高了 61% (p=0.060, N.S.）比 MMP 合成率高。 这些蛋白质的分离 通过 GC/C/IRMS 进行同位素丰度分析提供了有价值的信息 有关这些特定收缩的合成速率的信息 蛋白质，而不是提供的更一般的信息 先前对 MMP 合成速率的研究。