Integrated Dissection of Cysteine Oxidative Post-translational Modification Proteome During Cardiac Hypertrophy

Abstract

Cysteine oxidative modification of cellular proteins is crucial for many aspects of cardiac hypertrophy development. However, integrated dissection of multiple types of cysteine oxidative post-translational modifications (O-PTM) of proteomes in cardiac hypertrophy is currently missing. Here we developed a novel discovery platform that encompasses a customized biotin switch-based quantitative proteomics pipeline and an advanced analytic workflow to comprehensively profile the landscape of cysteine O-PTM in an ISO-induced cardiac hypertrophy mouse model. Specifically, we identified a total of 1655 proteins containing 3324 oxidized cysteine sites by at least one of the following three modifications: reversible cysteine O-PTM, cysteine sulfinylation (CysSOH), and cysteine sulfonylation (CysSOH). Analyzing the hypertrophy signatures that are reproducibly discovered from this computational workflow unveiled four biological processes with increased cysteine O-PTM. Among them, protein phosphorylation, creatine metabolism, and response to elevated Ca pathways exhibited an elevation of cysteine O-PTM in early stages, whereas glucose metabolism enzymes were increasingly modified in later stages, illustrating a temporal regulatory map in cardiac hypertrophy. Our cysteine O-PTM platform depicts a dynamic and integrated landscape of the cysteine oxidative proteome, through the extracted molecular signatures, and provides critical mechanistic insights in cardiac hypertrophy. Data are available via ProteomeXchange with identifier PXD010336.

Toward Completion of the Human Proteome Parts List: Progress Uncovering Proteins That Are Missing or Have Unknown Function and Developing Analytical Methods

“…Wang et al 36 provide a deep analysis of the cysteine oxidative proteome in cardiac hypertrophy, a key feature of hypertensive heart disease, congestive heart failure, and sudden cardiac death. Reactive oxygen and nitrogen species (RS/NS) mediate hypertrophy through multiple types of oxidation of protein cysteine residues, triggering molecular switches.…”

Irreversible oxidative post-translational modifications in heart disease

“…A combination of independent established markers as suggested early on by Sabatine et al [108] can improve the coverage of different types of heart failure but does not alleviate the problem of specific detection of earlystage disease. Promising new biomarkers based on oxidative post-translational modifications of cardiac proteins have been reported by several independent research groups [109][110][111][112]. A direct relationship of oxidative modifications of proteins and cardiac function was shown in murine animal models [113].…”

A high-stringency blueprint of the human proteome

“…Over the decade, CVD proteomics has broadened from identifying single canonical proteins to mapping proteoforms derived from combinations of alternative splicing, cleavage, and PTMs 33 , 83 . Now, identification of proteoforms (e.g., genetic variations, alternatively spliced products, phosphorylation 118 , glycosylation 119 , oxidative 120 and other PTMs 121 ) allow CVD sub-classification. In parallel, the heart is uniquely sensitive to alternative splicing (frequently altered in congenital heart diseases), explaining the B/D-HPP’s interest in developing assays for splice isoform-specific changes in cardiomyocyte development and maturation 122 , 123 .…”