Proteomic study of the mechanism of talin-C as an inhibitor of HIV infection

IF 0.5 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Current Proteomics Pub Date : 2020-12-14 DOI:10.2174/1570164618999201214153239

L. Yin, Yujiao Zhang, Huichun Shi, Ya-ru Xing, Hong Zhou Lu, Lijun Zhang

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Abstract

Talin-1 is involved in human immunodeficiency virus (HIV) invasion and synapse development. We found that talin-1 was cleaved into a 38 KDa fragment (talin-C) in the peripheral blood mononuclear cells (PBMCs) of HIV patients; however, the underlying mechanisms remain unknown. This study aimed to determine the relationship between talin-C and HIV infection and identify the mechanisms underlying the ability of this protein to influence HIV infection. PBMCs were derived from HIV-infected patients enrolled in this study. N- and C-terminal peptides matching the potential sequence of talin-C were detected in PBMCs by multiple reaction monitoring (MRM) mass spectrometry. TZM-b1 cells were infected with HIV-1 pseudotyped virus (HIVpp) for different durations to detect the talin-C product. Three stable cell lines overexpressing talin head (TLN1-H) or TLN1-C or with TLN1 knockdown (shTLN1) were created and infected by HIVpp. The HIV marker protein (P24) was then detected by enzyme-linked immunosorbent assay. Finally, an isobaric tag for relative and absolute quantification (iTRAQ)-based proteomic study was performed to detect the TLN1-C-regulated proteins with or without HIVpp infection in TZM-bl cells. The identified proteins were analyzed by R version 4.0.2, and STRING software (Version: 11.0) (https://string-db.org). N- and C-peptides of talin-C were detected to have higher expression in patients with lower HIV load. Talin-C was produced during HIVpp infection. TLN1-C significantly inhibited HIVpp infection in the TZM-b1 cells. Additionally, a proteomic study found that TLN1-C regulated the expression of 99 proteins in TZM-b1 cells without and with HIVpp infection, respectively. According to Gene Ontology (GO) annotation, proteins with cellular metabolic process and binding function were found to be enriched. Thirty four proteins have protein-protein interaction, including 19 down- and 15 up- regulated proteins, respectively. Talin-C was produced following HIV infection, and is inversely proportional to HIV load. A proteomic study indicated that TLN1-C might be involved in HIV infection through regulating metabolic processes.

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talin-C作为HIV感染抑制剂的蛋白质组学研究

Talin-1参与人类免疫缺陷病毒(HIV)侵袭和突触发育。我们发现talin-1在hiv患者外周血单核细胞(PBMCs)中被切割成一个38 KDa的片段(talin-C);然而，其潜在机制尚不清楚。本研究旨在确定talin-C与HIV感染之间的关系，并确定该蛋白影响HIV感染的潜在机制。pbmc来源于参与本研究的hiv感染患者。采用多反应监测(MRM)质谱法检测了与talin-C潜在序列匹配的N端肽和c端肽。用HIV-1假型病毒(HIVpp)感染tzm -b1细胞不同时间，检测talin-C产物。建立了3株过表达talin head (TLN1- h)、TLN1- c或TLN1敲低(shTLN1)的稳定细胞系，并用hivpp感染。然后用酶联免疫吸附法检测HIV标记蛋白(P24)。最后，采用等压标记相对和绝对定量(iTRAQ)方法进行蛋白质组学研究，检测感染或未感染HIVpp的TZM-bl细胞中tln1 - c调节蛋白。用R 4.0.2版本和string软件(版本:11.0)(https://string-db.org).N-)对鉴定的蛋白进行分析，检测talin-C的c肽在HIV载量较低的患者中表达较高。Talin-C是在hiv感染期间产生的。TLN1-C显著抑制TZM-b1细胞的HIVpp感染。此外，一项蛋白质组学研究发现，TLN1-C分别调节未感染和感染HIVpp的TZM-b1细胞中99种蛋白的表达。根据基因本体(Gene Ontology, GO)注释，发现具有细胞代谢过程和结合功能的蛋白质富集。34种蛋白质之间有相互作用，分别包括19种下调蛋白和15种上调蛋白。Talin-C是在HIV感染后产生的，与HIV载量成反比。一项蛋白质组学研究表明，TLN1-C可能通过调节代谢过程参与HIV感染。

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来源期刊

Current Proteomics BIOCHEMICAL RESEARCH METHODS-BIOCHEMISTRY & MOLECULAR BIOLOGY

CiteScore

1.60

自引率

0.00%

发文量

审稿时长

>0 weeks

期刊介绍： Research in the emerging field of proteomics is growing at an extremely rapid rate. The principal aim of Current Proteomics is to publish well-timed in-depth/mini review articles in this fast-expanding area on topics relevant and significant to the development of proteomics. Current Proteomics is an essential journal for everyone involved in proteomics and related fields in both academia and industry. Current Proteomics publishes in-depth/mini review articles in all aspects of the fast-expanding field of proteomics. All areas of proteomics are covered together with the methodology, software, databases, technological advances and applications of proteomics, including functional proteomics. Diverse technologies covered include but are not limited to: Protein separation and characterization techniques 2-D gel electrophoresis and image analysis Techniques for protein expression profiling including mass spectrometry-based methods and algorithms for correlative database searching Determination of co-translational and post- translational modification of proteins Protein/peptide microarrays Biomolecular interaction analysis Analysis of protein complexes Yeast two-hybrid projects Protein-protein interaction (protein interactome) pathways and cell signaling networks Systems biology Proteome informatics (bioinformatics) Knowledge integration and management tools High-throughput protein structural studies (using mass spectrometry, nuclear magnetic resonance and X-ray crystallography) High-throughput computational methods for protein 3-D structure as well as function determination Robotics, nanotechnology, and microfluidics.