Background: Human seminal prostasomes are intrinsically heterogeneous extracellular vesicles (EVs) whose composition is, additionally, influenced by different physiological conditions. Aiming at the molecular properties of the prostasomal surface exemplified by glycan compositions as a possible distinction factor, we applied lectin-affinity chromatography (LAC) as a new tool for their separation. Since glycans, generally, exhibit various biological activities, introduction of glyco-parameters as reference could upgrade standardization of EVs isolated by different methods and intended for use in biomedicine.Methods: Preparations of seminal prostasomes from normozoospermic (sPro-N) and oligozoospermic (sPro-O) men were subjected to LAC on concanavalin A (Con A) and wheat germ agglutinin (WGA) columns. Prostasomes recovered in LAC-separated fractions were characterized according to the distribution of selected markers: gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), tetraspanin CD63, and total protein/glycoprotein composition.Results: Two CD63-immunoreactive populations exhibiting prostasome signature bands but differing in GGT activity and surface glycans were separated on the WGA column. Additional populations having distinct profiles of total glycoproteins and which can be tracked down by ALP activity were enriched on the Con A column. WGA-separated populations were similar in sPro-N and sPro-O, whereas Con A-separated ones were strikingly different.Conclusions: Membrane-associated gamma-glutamyl transferase and alkaline phosphatase in the context of Con A- and WGA-reactive glycans mark seminal prostasomes populations from normozoospermic and oligozoospermic men.

中文翻译：

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在伴刀豆球蛋白A和小麦胚芽凝集素反应性聚糖的背景下，膜相关的γ-谷氨酰转移酶和碱性磷酸酶标记来自正常精子和少精子症男性的精囊。

背景：人类精原体是本质上异质的细胞外囊泡（EVs），其组成还受到不同生理条件的影响。针对以聚糖组合物为例的前体表面的分子特性作为可能的区分因素，我们将凝集素亲和色谱法（LAC）用作分离它们的新工具。由于聚糖通常表现出多种生物学活性，因此引入糖参数作为参考可以提高通过不同方法分离并打算用于生物医学的电动汽车的标准化方法。 -O）男子在伴刀豆球蛋白A（Con A）和小麦胚芽凝集素（WGA）柱上进行了LAC。根据所选标记的分布表征在LAC分离的级分中回收的前列腺小体：γ-谷氨酰转移酶（GGT），碱性磷酸酶（ALP），四跨素CD63和总蛋白/糖蛋白组成。结果：两个具有前列腺小体的CD63免疫反应人群在WGA色谱柱上分离出了一些特征性条带，但它们的GGT活性和表面聚糖不同。在Con A色谱柱上富集了具有不同总糖蛋白特征并可以通过ALP活性追踪的其他种群。WGA分离的群体在sPro-N和sPro-O中相似，而用Con A分离的群体则截然不同。