de Mateo Sara, Estanyol Josep Maria, Oliva Rafael
Human Genetics Research Group, IDIBAPS, University of Barcelona, and Biochemistry and Molecular Genetics Service, Hospital Clínic i Provincial, Barcelona, Spain.
Methods Mol Biol. 2013;927:411-22. doi: 10.1007/978-1-62703-038-0_35.
Proteomics is the study of the proteins of cells or tissues. Sperm proteomics aims at the identification of the proteins that compose the sperm cell and the study of their function. The recent developments in mass spectrometry (MS) have markedly increased the throughput for the identification and study of the sperm proteins. Catalogues of spermatozoal proteins in human and in model species are becoming available laying the groundwork for subsequent research, diagnostic applications, and the development of patient-specific treatments. A wide range of MS techniques is also rapidly becoming available for researchers. This chapter describes a methodological option to study the sperm cell using MS and provides a detailed protocol to identify the proteins extracted from a Percoll-purified human sperm population and separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) using LC-MS/MS.
蛋白质组学是对细胞或组织中的蛋白质进行的研究。精子蛋白质组学旨在鉴定构成精子细胞的蛋白质并研究其功能。质谱(MS)技术的最新进展显著提高了精子蛋白质鉴定和研究的通量。人类和模式物种的精子蛋白质目录正逐渐可得,为后续研究、诊断应用以及个性化治疗的开发奠定了基础。各种质谱技术也迅速可供研究人员使用。本章描述了一种使用质谱研究精子细胞的方法选择,并提供了一份详细方案,用于鉴定从经Percoll纯化的人类精子群体中提取并通过二维聚丙烯酰胺凝胶电泳(2D-PAGE)分离的蛋白质,随后使用液相色谱-质谱联用(LC-MS/MS)进行分析。