Yin Ping, Hou Xiaowen, Romanova Elena V, Sweedler Jonathan V
Department of Chemistry, University of Illinois, Urbana, IL, USA.
Methods Mol Biol. 2011;789:223-36. doi: 10.1007/978-1-61779-310-3_14.
Neuropeptidomics refers to a global characterization approach for the investigation of neuropeptides, often under specific physiological conditions. Neuropeptides comprise a complex set of signaling molecules that are involved in regulatory functions and behavioral control in the nervous system. Neuropeptidomics is inherently challenging because neuropeptides are spatially, temporally, and chemically heterogeneous, making them difficult to predict in silico from genomic information. Mature neuropeptides are produced from intricate enzymatic processing of precursor proteins/prohormones via a range of posttranslational modifications, resulting in multiple final peptide products from each prohormone gene. Although there are several methods for targeted peptide studies, mass spectrometry (MS), with its qualitative and quantitative capabilities, is ideally suited to the task. MS provides fast, sensitive, accurate, and high-throughput peptidomic analysis of neuropeptides without requiring prior knowledge of the peptide sequences. Aided by liquid chromatography (LC) separations and bioinformatics, MS is quickly becoming a leading technique in neuropeptidomics. This chapter describes several LC-MS analytical methods to identify, characterize, and quantify neuropeptides while emphasizing the sample preparation steps so integral to experimental success.
神经肽组学是一种用于研究神经肽的全局表征方法，通常是在特定生理条件下进行研究。神经肽是一组复杂的信号分子，参与神经系统的调节功能和行为控制。神经肽组学本身具有挑战性，因为神经肽在空间、时间和化学性质上具有异质性，这使得难以从基因组信息中通过计算机模拟进行预测。成熟的神经肽是通过前体蛋白/前激素的复杂酶促加工过程，并经过一系列翻译后修饰产生的，每个前激素基因会产生多种最终肽产物。虽然有几种针对特定肽的研究方法，但质谱（MS）凭借其定性和定量能力，非常适合这项任务。质谱无需事先了解肽序列，就能对神经肽进行快速、灵敏、准确且高通量的肽组学分析。借助液相色谱（LC）分离和生物信息学，质谱正迅速成为神经肽组学的主导技术。本章介绍了几种用于鉴定、表征和定量神经肽的液相色谱 - 质谱分析方法，同时强调了对于实验成功至关重要的样品制备步骤。
