2026-04-14T03:37:10Zhttps://recercat.cat/oai/request

oai:recercat.cat:2445/2180712025-12-04T19:09:39Zcom_2072_1057col_2072_478780col_2072_478858col_2072_478917

00925njm 22002777a 4500 dc Soler Ventura, Ada author Gay i Marín, Marina author Jodar Bifet, Meritxell author Vilanova, Mar author Castillo Corullón, Judit author Arauz-Garofalo, Gianluca author Villarreal, Laura author Ballescà, Josep Lluís author Vilaseca, Marta author Oliva Virgili, Rafael author 2025-01-28T14:47:41Z 2025-01-28T14:47:41Z 2020-01-03 2025-01-28T14:47:41Z Protamine 1 (P1) and protamine 2 (P2) family are extremely basic, sperm-specific proteins, packing 85-95% of the paternal DNA. P1 is synthesized as a mature form, whereas P2 components (HP2, HP3, and HP4) arise from the proteolysis of the precursor (pre-P2). Due to the particular protamine physical-chemical properties, their identification by standardized bottom-up mass spectrometry (MS) strategies is not straightforward. Therefore, the aim of this study was to identify the sperm protamine proteoforms profile, including their post-translational modifications, in normozoospermic individuals using two complementary strategies, a top-down MS approach and a proteinase-K-digestion-based bottom-up MS approach. By top-down MS, described and novel truncated P1 and pre-P2 proteoforms were identified. Intact P1, pre-P2, and P2 mature proteoforms and their phosphorylation pattern were also detected. Additionally, a +61 Da modification in different proteoforms was observed. By the bottom-up MS approach, phosphorylated residues for pre-P2, as well as the new P2 isoform 2, which is not annotated in the UniProtKB database, were revealed. Implementing these strategies in comparative studies of different infertile phenotypes, together with the evaluation of P1/P2 and pre-P2/P2 MS-derived ratios, would permit determining specific alterations in the protamine proteoforms and elucidate the role of phosphorylation/dephosphorylation dynamics in male fertility. Espermatozoides Fosforilació Esterilitat masculina Protamines Proteòmica Spermatozoa Phosphorylation Male sterility Protamines Proteomics Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches