Human skeletal muscle fiber type specific protein content
- 作者：Andrew J. Galpin ; Ulrika Raue ; Bozena Jemiolo ; Todd A. Trappe ; Matthew P. Harber ; Kiril Minchev ; Scott Trappe ; strappe@bsu.edu
- 关键词：Western blot ; GAPDH ; Citrate synthase ; p38
- 刊名：Analytical Biochemistry
- 出版年：2012
- 期刊代码：93_00032697
- 类别：imm
- 出版时间：15 June, 2012
- 卷：425
- 期：2
- 页码：175-182
- 文件大小：532 K
摘要
The aim of this project was to develop a method to assess fiber type specific protein content across the continuum of human skeletal muscle fibers. Individual vastus lateralis muscle fibers (n = 264) were clipped into two portions: one for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) fiber typing and one for Western blot protein identification. Following fiber type determination, fiber segments were combined into fiber type specific pools (鈭?20 fibers/pool) and measured for total protein quantity, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), citrate synthase (CS), and total p38 content. GAPDH content was 64, 54, 160, and 138%more abundant in myosin heavy chain (MHC) I/IIa, MHC IIa, MHC IIa/IIx, and MHC IIx fibers, respectively, when compared with MHC I. Inversely, CS content was 528, 472, 242, and 47%more abundant in MHC I, MHC I/IIa, MHC IIa, and MHC IIa/IIx fibers, respectively, when compared with MHC IIx. Total p38 content was 87%greater in MHC IIa versus MHC I fibers. These data and this approach establish a reliable method for human skeletal muscle fiber type specific protein analysis. Initial results show that particular proteins exist in a hierarchal fashion throughout the continuum of human skeletal muscle fiber types, further highlighting the necessity of fiber type specific analysis.