Enrichment of Cysteine-Containing Peptide by On-Resin Capturing and Fixed Charge Tag Derivatization for Sensitive ESI-MS Detection

Molecules

Karolina Panek-Laszczyńska¹  Karolina Mojsa²  Remigiusz Bąchor³  Zbigniew Szewczuk³  Oliwia Gorzeń³  Anna Rola³  Krystyna Dąbrowska⁴  Wojciech Witkiewicz⁵  Andrzej Konieczny⁶ 

[1] 1st Department and Clinic of Gynecology and Obstetrics, Wroclaw Medical University, 50-368 Wroclaw, Poland;Department of Molecular Microbiology, Faculty of Biotechnology, University of Wroclaw, 50-383 Wroclaw, Poland;Faculty of Chemistry, University of Wroclaw, 50-383 Wroclaw, Poland;Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 53-114 Wroclaw, Poland;Research and Development Center, Regional Specialized Hospital, 51-124 Wroclaw, Poland;Wroclaw Medical University, Department of Nephrology and Transplantation Medicine, 50-556 Wroclaw, Poland;

关键词: cysteine;    thiopeptides;    maleimide;    mass spectrometry;    fixed charge tags;    podocin;   

DOI  :  10.3390/molecules25061372

来源: DOAJ

【 摘 要 】

High complexity of cell and tissue proteomes limits the investigation of proteomic biomarkers. Therefore, the methods of enrichment of some chemical groups of peptides including thiopeptides are important tools that may facilitate the proteomic analysis by reducing sample complexity and increasing proteome coverage. Here, we present a new method of cysteine-containing tryptic peptide enrichment using commercially available TentaGel R RAM resin modified by the linker containing the maleimide group, allowing thiol conjugation. The captured tryptic peptides containing lysine residue were then tagged by 2,4,6-triphenylpyrylium salt to form 2,4,6-triphenylpyridinium derivatives, which increases the ionization efficiency during mass spectrometry analysis. This makes it possible to conduct an ultrasensitive analysis of the trace amount of compounds. The proposed strategy was successfully applied in the enrichment of model tryptic podocin peptide and podocin tryptic digest.

【 授权许可】

Unknown