The Strategy for Peptidomic LC-MS/MS Data Analysis: The Case of Urinary Peptidome Study

Methods Mol Biol. 2024:2758:389-399. doi: 10.1007/978-1-0716-3646-6_21.

Abstract

The study of urinary peptidome is an important area of research, which concerns the characterization of endogenous peptides, as well as the identification of biomarkers for a wide range of socially significant diseases. First of all, this relates to renal and genitourinary pathologies and/or pathologies associated with proteinuria, such as kidney diseases, bladder, prostate and ovarian cancers, diabetic nephropathy, and pre-eclampsia. Unlike proteins, peptides do not require proteolytic hydrolysis, can be analyzed in their native form and can provide certain information about occurring (patho)physiological processes. Mass spectrometry (MS)-based approaches are the most unbiased and sensitive instruments with high multiplexing capacity and provided most of the current information about endogenous urine peptides. However, despite the large number of urine peptidomic studies, there are certain issues related to the insufficient comparability of their results due to the lack of consistent approaches to their interpretation. Also the development of a custom project-specific protein library for endogenous peptides search and identification is another important point that should be noted in the context of high-throughput peptidomic analysis. Here we propose the custom-specific urinary protein database and the grouping of endogenous urinary peptides with overlapping sequences as useful tools, which can facilitate the acquisition and analysis of LC-MS peptidomic data, as well as the comparison of results of different studies, which should facilitate their more efficient further application.

Keywords: Endogenous peptides; Mass spectrometry; Pre-eclampsia; Proteinuria; Sequence alignment; Urinary protein database; Urine peptidome.

MeSH terms

  • Chromatography, Liquid
  • Female
  • Humans
  • Liquid Chromatography-Mass Spectrometry*
  • Male
  • Peptides / metabolism
  • Pregnancy
  • Proteins
  • Proteomics / methods
  • Tandem Mass Spectrometry* / methods

Substances

  • Proteins
  • Peptides