Protamine Characterization by Top-Down Proteomics: Boosting Proteoform Identification with DBSCAN

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ID: 268118
2021
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Abstract
Protamines replace histones as the main nuclear protein in the sperm cells of many species and play a crucial role in compacting the paternal genome. Human spermatozoa contain protamine 1 (P1) and the family of protamine 2 (P2) proteins. Alterations in protamine PTMs or the P1/P2 ratio may be associated with male infertility. Top-down proteomics enables large-scale analysis of intact proteoforms derived from alternative splicing, missense or nonsense genetic variants or PTMs. In contrast to current gold standard techniques, top-down proteomics permits a more in-depth analysis of protamine PTMs and proteoforms, thereby opening up new perspectives to unravel their impact on male fertility. We report on the analysis of two normozoospermic semen samples by top-down proteomics. We discuss the difficulties encountered with the data analysis and propose solutions as this step is one of the current bottlenecks in top-down proteomics with the bioinformatics tools currently available. Our strategy for the data analysis combines two software packages, ProSight PD (PS) and TopPIC suite (TP), with a clustering algorithm to decipher protamine proteoforms. We identified up to 32 protamine proteoforms at different levels of characterization. This in-depth analysis of the protamine proteoform landscape of normozoospermic individuals represents the first step towards the future study of sperm pathological conditions opening up the potential personalized diagnosis of male infertility.
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arauz-garofalo2021proteomesprotamine Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Gianluca Arauz-Garofalo;Meritxell Jodar;Mar Vilanova;Alberto de la Iglesia Rodriguez;Judit Castillo;Ada Soler-Ventura;Rafael Oliva;Marta Vilaseca;Marina Gay;Arauz-Garofalo, Gianluca;Jodar, Meritxell;Vilanova, Mar;de la Iglesia Rodriguez, Alberto;Castillo, Judit;Soler-Ventura, Ada;Oliva, Rafael;Vilaseca, Marta;Gay, Marina;
Journal proteomes
Year 2021
DOI 10.3390/proteomes9020021
URL
https://doi.org/10.3390/proteomes9020021
Keywords
male infertility bioinformatics lc-ms/ms sperm post-translational modifications dbscan protamine top-down proteomics proteoform

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