The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection.
Insertion of T4 lysozyme (T4L) into the GPCR successfully enhanced GPCR protein stability and solubilization. However, the biological functions of the recombinant GPCR protein have not been analyzed.We engineered the CCR5-T4L mutant and expressed and purified the soluble recombinant protein using an...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4496087?pdf=render |
| _version_ | 1818236657949736960 |
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| author | Qingwen Jin Hong Chen Xingxia Wang Liandong Zhao Qingchen Xu Huijuan Wang Guanyu Li Xiaofan Yang Hongming Ma Haoquan Wu Xiaohui Ji |
| author_facet | Qingwen Jin Hong Chen Xingxia Wang Liandong Zhao Qingchen Xu Huijuan Wang Guanyu Li Xiaofan Yang Hongming Ma Haoquan Wu Xiaohui Ji |
| author_sort | Qingwen Jin |
| collection | DOAJ |
| description | Insertion of T4 lysozyme (T4L) into the GPCR successfully enhanced GPCR protein stability and solubilization. However, the biological functions of the recombinant GPCR protein have not been analyzed.We engineered the CCR5-T4L mutant and expressed and purified the soluble recombinant protein using an E.coli expression system. The antiviral effects of this recombinant protein in THP-1 cell lines, primary human macrophages, and PBMCs from different donors were investigated. We also explored the possible mechanisms underlying the observed antiviral effects.Our data showed the biphasic inhibitory and promotion effects of different concentrations of soluble recombinant CCR5-T4L protein on R5 tropic human immunodeficiency virus-1 (HIV-1) infection in THP-1 cell lines, human macrophages, and PBMCs from clinical isolates. We demonstrated that soluble recombinant CCR5-T4L acts as a HIV-1 co-receptor, interacts with wild type CCR5, down-regulates the surface CCR5 expression in human macrophages, and interacts with CCL5 to inhibit macrophage migration. Using binding assays, we further determined that recombinant CCR5-T4L and [125I]-CCL5 compete for the same binding site on wild type CCR5.Our results suggest that recombinant CCR5-T4L protein marginally promotes HIV-1 infection at low concentrations and markedly inhibits infection at higher concentrations. This recombinant protein may be helpful in the future development of anti-HIV-1 therapeutic agents. |
| first_indexed | 2024-12-12T12:13:21Z |
| format | Article |
| id | doaj.art-0c9906c3c75a4fcf82673df040ab0cc7 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T12:13:21Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-0c9906c3c75a4fcf82673df040ab0cc72022-12-22T00:24:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-01107e013189410.1371/journal.pone.0131894The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection.Qingwen JinHong ChenXingxia WangLiandong ZhaoQingchen XuHuijuan WangGuanyu LiXiaofan YangHongming MaHaoquan WuXiaohui JiInsertion of T4 lysozyme (T4L) into the GPCR successfully enhanced GPCR protein stability and solubilization. However, the biological functions of the recombinant GPCR protein have not been analyzed.We engineered the CCR5-T4L mutant and expressed and purified the soluble recombinant protein using an E.coli expression system. The antiviral effects of this recombinant protein in THP-1 cell lines, primary human macrophages, and PBMCs from different donors were investigated. We also explored the possible mechanisms underlying the observed antiviral effects.Our data showed the biphasic inhibitory and promotion effects of different concentrations of soluble recombinant CCR5-T4L protein on R5 tropic human immunodeficiency virus-1 (HIV-1) infection in THP-1 cell lines, human macrophages, and PBMCs from clinical isolates. We demonstrated that soluble recombinant CCR5-T4L acts as a HIV-1 co-receptor, interacts with wild type CCR5, down-regulates the surface CCR5 expression in human macrophages, and interacts with CCL5 to inhibit macrophage migration. Using binding assays, we further determined that recombinant CCR5-T4L and [125I]-CCL5 compete for the same binding site on wild type CCR5.Our results suggest that recombinant CCR5-T4L protein marginally promotes HIV-1 infection at low concentrations and markedly inhibits infection at higher concentrations. This recombinant protein may be helpful in the future development of anti-HIV-1 therapeutic agents.http://europepmc.org/articles/PMC4496087?pdf=render |
| spellingShingle | Qingwen Jin Hong Chen Xingxia Wang Liandong Zhao Qingchen Xu Huijuan Wang Guanyu Li Xiaofan Yang Hongming Ma Haoquan Wu Xiaohui Ji The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection. PLoS ONE |
| title | The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection. |
| title_full | The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection. |
| title_fullStr | The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection. |
| title_full_unstemmed | The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection. |
| title_short | The Effects of the Recombinant CCR5 T4 Lysozyme Fusion Protein on HIV-1 Infection. |
| title_sort | effects of the recombinant ccr5 t4 lysozyme fusion protein on hiv 1 infection |
| url | http://europepmc.org/articles/PMC4496087?pdf=render |
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