University of Belgrade, Institute for the Application of Nuclear Energy, INEP, Belgrade, Serbia.
University of Belgrade, Institute of Nuclear Sciences, VINČA, National Institute of the Republic of Serbia, Belgrade, Serbia.
Ups J Med Sci. 2021 Sep 3;126. doi: 10.48101/ujms.v126.7673. eCollection 2021.
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.
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.
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.
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.
前列腺小体是存在于精液中的细胞外囊泡（EVs），大量表达独特的四跨膜蛋白（TS）和半乳糖凝集素-3（gal-3），它们通过不同分子复合物的组装来塑造其表面。在这项研究中，我们确定了膜相关前列腺小体蛋白的去污剂敏感性模式，旨在确定其作为内在多分子标记的可溶性特征，以及作为健康和疾病中 EVs 群体比较的参考新参数。
用 Triton X-100 破坏前列腺小体，并在保持完全可溶性的条件下通过凝胶过滤进行分析。使用固相凝集素结合测定、透射电子显微镜、电泳和凝集素印迹监测 TS（CD63、CD9 和 CD81）、gal-3、γ-谷氨酰转移酶（GGT）和不同 N-聚糖的再分布。
关于正常生理和精子计数低的条件下前列腺小体的比较数据表明，不同 N-聚糖和 GGT 的再分布具有相似性，所有这些都被认为主要是囊泡外壳的一部分。与此相反，在整合膜蛋白的再分布中发现了更大的差异，以 TS 和 gal-3 为例。因此，它们分为两种主要由重叠的 CD9/gal-3/麦胚凝集素反应性糖蛋白和 CD63/GGT/伴刀豆球蛋白 A 反应性糖蛋白组成的分子模式。
由于它反映了亲代细胞和细胞外环境的状态，因此可溶性特征可以被认为是特定囊泡表面特性的一个综合区别因素，这两者都有助于空间膜排列的组成。
