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(肌)营养不良短小蛋白结合蛋白1(dystrobrevin binding protein 1,dysbindin-1)是溶酶体相关细胞器生物发生复合体-1(biogenesis of lysosome-related organelles complex 1,BLOC-1)的1个亚基,在多种组织细胞中广泛表达;然而,其在睾丸组织中的作用至今尚不明确。为寻找(肌)营养不良短小蛋白结合蛋白1在睾丸组织中的相互作用蛋白质,以进一步研究(肌)营养不良短小蛋白结合蛋白1在睾丸中的作用,本研究首先在Rosetta(DE3)菌种中表达可溶性GST-dysbindin-1融合蛋白,经谷胱甘肽-琼脂糖珠亲和纯化后,与小鼠的睾丸组织蛋白质孵育进行GST pull-down实验,并通过液相色谱串联质谱(LC MS/MS)分析筛选(肌)营养不良短小蛋白结合蛋白1在睾丸组织中的相互作用蛋白质。利用Bio GPS数据库聚类在睾丸组织中高表达和特异性表达的互作蛋白质,运用DAVID6.8在线分析工具从细胞组分、分子功能、生物学过程和KEGG通路等方面对筛选出的互作蛋白质进行GO(gene ontology)富集分析。本实验共筛选出108个(肌)营养不良短小蛋白结合蛋白1在睾丸组织中的潜在互作蛋白质,其中98个为尚未报道的(肌)营养不良短小蛋白结合蛋白1相互作用蛋白质,7个为睾丸高表达蛋白质,5个为睾丸特异性表达的蛋白质。这些候选蛋白质主要分布在细胞质、细胞核、细胞膜、细胞外泌体等细胞组分中,通过与蛋白质、核酸等分子结合参与蛋白质翻译和转运、囊泡运输及凋亡等生物学过程以及氨基酸生物合成、溶酶体及蛋白酶体等生物学通路。我们推测,在睾丸组织中(肌)营养不良短小蛋白结合蛋白1可能通过与多种蛋白质相互作用参与精子的发生和受精等过程。
Dystrobrevin binding protein 1 (dysbindin-1) is one subfamily of biogenesis of lysosome-related organelles complex 1 (BLOC-1) It is widely expressed in a variety of tissue cells; however, its role in testicular tissue remains unclear. In order to investigate the role of dystrophin short protein-binding protein-1 in the testis, we investigated the interaction between malnourished short protein-binding protein 1 and testis in Rosetta (DE3) GST pull-down assay was performed on the mouse testis tissue proteins after affinity purification with glutathione-agarose beads and by liquid chromatography tandem mass spectrometry (LC MS) / MS) Analysis of Screening (Muscular) Malnutrition The interaction of short protein-binding protein 1 in testis tissue proteins. Using BioGDB database to cluster the high expression and specific expression of interaction protein in testis tissue, using the on-line analysis tool DAVID6.8 from the cell components, molecular function, biological processes and KEGG pathway screening of the interaction protein GO (gene ontology) enrichment analysis. In this study, we identified a total of 108 potential proteins interacting in the testis of 108 (muscle) malnourished short protein-binding protein 1s, of which 98 were unreported (dystrophic short protein-binding protein-1 interacting proteins, 7 High expression of testicular protein, 5 for testicular specific expression of protein. These candidate proteins are mainly distributed in cell components such as cytoplasm, nucleus, cell membrane and exocytosis, and are involved in the biological processes such as protein translation and transport, vesicle trafficking and apoptosis, and amino acid biosynthesis through binding to proteins and nucleic acids , Lysosomes and proteasome and other biological pathways. We speculate that malnutrition in the testicular tissue short protein-binding protein 1 may participate in the process of spermatogenesis and fertilization by interacting with various proteins.