Automated Phosphopeptide Enrichment for Gram-Positive Bacteria.

ABSTRACT: Protein phosphorylation in prokaryotes has gained more attention in recent years as several studies linked it to regulatory and signaling functions, indicating importance similar to protein phosphorylation in eukaryotes. Studies on bacterial phosphorylation have so far been conducted using manual or HPLC-supported phosphopeptide enrichment, whereas automation of phosphopeptide enrichment has been established in eukaryotes, allowing for high-throughput sampling. To facilitate the prospect of studying bacterial phosphorylation on a systems level, we here established an automated Ser/Thr/Tyr phosphopeptide enrichment workflow on the Agilent AssayMap platform. We present optimized buffer conditions for TiO₂ and Fe(III)-NTA-IMAC cartridge-based enrichment and the most advantageous, s

SUBMITTER: Birk MS 

PROVIDER: S-EPMC8491273 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature