Lin Yin, Yujiao Zhang, Huichun Shi, Yaru Xing, Hongzhou Lu and Lijun Zhang* Pages 705 - 716 ( 12 )
Background: Talin-1 is involved in the invasion and synapse development of the Human Immunodeficiency Virus (HIV). 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.
Objective: This study aimed to determine the relationship between talin-C and HIV infection and to identify the mechanisms underlying the ability of this protein to influence HIV infection.
Methods: 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 the 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 proteomics 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).
Results: 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 with and without HIVpp infection, respectively. According to Gene Ontology (GO) annotation, proteins with cellular metabolic processes and binding function were found to be enriched. Thirty-four proteins have protein-protein interaction, 19 down- and 15 up-regulated proteins, respectively.
Conclusion: Talin-C was produced following HIV infection and was inversely proportional to HIV load. A proteomic study indicated that TLN1-C might be involved in HIV infection through regulating metabolic processes.
Talin-C, HIV, proteomics, iTRAQ, metabolic pathways, STRING software.
Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508