一种适应性基因组-蛋白质组学方法表征细胞表面Vimentin的膜拓扑结构和相互作用

IF 2.2 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2025-09-06 DOI:10.1016/j.bbrep.2025.102242

Masoud Norouzi, Christopher A. McCulloch

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引用次数: 0

摘要

当在细胞表面发现Vimentin时，它是一种细胞骨架蛋白，参与多种病理生理过程。在人类成纤维细胞和内皮细胞中，我们使用CRISPR-Cas9在vim基因的六个不同位点插入血凝素标签。然后，我们利用标签免疫检测来定义细胞表面Vimentin （csVim）的膜拓扑结构，然后通过邻近生物素化来鉴定其细胞外相互作用组；用体外定量结合试验验证命中的一个子集。

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An adaptable genomic-proteomic approach to characterize cell-surface Vimentin's membrane topology and interactome

When found on the cell surface, Vimentin, a cytoskeletal protein, engages in multiple pathophysiological processes. Here, in human fibroblasts and endothelial cells, we used CRISPR-Cas9 to insert a hemagglutinin tag at six different loci within the vim gene. We then utilized tag immunodetection to define cell surface Vimentin's (csVim) membrane topology, followed by proximity biotinylation to identify its extracellular interactome; validating a subset of hits with a quantitative in vitro binding assay.

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.