o:25139 Assembly of tetraspanins, galectin-3, and distinct N-glycans defines the solubilization signature of seminal prostasomes from normozoospermic and oligozoospermic men en ABSTRACT Background: Prostasomes, extracellular vesicles (EVs) abundantly present in seminal plasma, express distinct tetraspanins (TS) and galectin-3 (gal-3), which are supposed to shape their surface by an assembly of different molecular complexes. In this study, detergent-sensitivity patterns of membrane-associated prostasomal proteins were determined aiming at the solubilization signature as an intrinsic multimolecular marker and a new parameter suitable as a reference for the comparison of EVs populations in health and disease. Methods: Prostasomes were disrupted by Triton X-100 and analyzed by gel filtration under conditions that maintained complete solubilization. Redistribution of TS (CD63, CD9, and CD81), gal-3, gamma-glutamyltransferase (GGT), and distinct N-glycans was monitored using solid-phase lectin-binding assays, transmission electron microscopy, electrophoresis, and lectin blot. Results: Comparative data on prostasomes under normal physiology and conditions of low sperm count revealed similarity regarding the redistribution of distinct N-glycans and GGT, all presumed to be mainly part of the vesicle coat. In contrast to this, a greater difference was found in the redistribution of integral membrane proteins, exemplified by TS and gal-3. Accordingly, they were grouped into two molecular patterns mainly consisting of overlapped CD9/gal-3/wheat germ agglutinin-reactive glycoproteins and CD63/ GGT/concanavalin A-reactive glycoproteins. Conclusions: Solubilization signature can be considered as an all-inclusive distinction factor regarding the surface properties of a particular vesicle since it reflects the status of the parent cell and the extracellular environment, both of which contribute to the composition of spatial membrane arrangements. extracellular vesicles, detergent sensitivity, CD63, CD9, gamma-glutamyl transferase, molecular patterns, normozoospermia, oligozoospermia 1552099 10.48101/ujms.v126.7673 2022-04-20T08:23:02.541Z 44 yes 46 Tamara Janković University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia 0000-0002-8188-0921 Jelena Danilović Luković University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor bioloških nauka person 0000-0003-2411-5032 Irena Miler University of Belgrade, Institute of Nuclear Sciences VINČA, Serbia person Ninoslav Mitić University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor nauka person Ljiljana Hajduković University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor nauka person Miroslava Janković University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor nauka person 0000-0002-7889-5110 application/pdf 2696173 https://phaidrabg.bg.ac.rs/o:25139 no yes 16 70 92000001 11A29 11A2903 11A28 Upsala journal of medical sciences 126 2021