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Integrated top-down and bottom-up proteomics mass spectrometry for the characterization of endogenous ribosomal protein heterogeneity

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Journal of Pharmaceutical Analysis 2023年第1期13卷 63-72页

作者： Ying Zhang Qinghua Cai Yuxiang Luo Yu Zhang Huilin Li School of Pharmaceutical Sciences Sun Yat-sen UniversityGuangzhou510006China Henan Engineering Laboratory for Mammary Bioreactor School of Life SciencesHenan UniversityKaifengHenan475004China The Shennong Laboratory Zhengzhou450002China Guangdong Key Laboratory of Chiral Molecule and Drug Discovery School of Pharmaceutical SciencesSun Yat-sen UniversityGuangzhou510006China

Ribosomes are abundant,large RNA-protein complexes that are the sites of all protein synthesis in *** in ribosomal proteins(RPs),including proteoforms arising from genetic variations,alternative splicing of RNA transcripts,post-translational modifications and alterations of protein expression level,have been linked to a diverse range of diseases,including cancer and *** characterization of ribosomal proteoforms is challenging but important for the discovery of potential disease biomarkers or protein *** the present work,using *** 70S RPs as an example,we first developed a top-down proteomics approach on a Waters Synapt G2 Si mass spectrometry(MS)system,and then applied it to the HeLa 80S *** results were complemented by a bottom-up *** total,50 out of 55 RPs were identified using the top-down *** these,more than 30 RPs were found to have their N-terminal methionine *** modifications such as methylation,acetylation,and hydroxylation were also observed,and the modification sites were identified by bottomup *** a HeLa 80S ribosomal sample,we identified 98 ribosomal proteoforms,among which multiple truncated 80S ribosomal proteoforms were observed,the type of information which is often overlooked by bottom-up *** their relevance to diseases is not yet known,the integration of topdown and bottom-up proteomics approaches paves the way for the discovery of proteoform-specific disease biomarkers or targets.

关键词： Ribosomal proteins Top-down MS Bottom-up MS proteoforms

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