Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors
Summary: It was posited that functionalities of GPCRs require full-length sequences that are negated by residue deletions. Here we report that significantly truncated nfCCR5QTY and nfCXCR4QTY still bind native ligands. Receptor-ligand interactions were discovered from yeast 2-hybrid screening and co...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-12-01
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| Series: | iScience |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004220308622 |
| _version_ | 1818393185852850176 |
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| author | Rui Qing Fei Tao Pranam Chatterjee Gaojie Yang Qiuyi Han Haeyoon Chung Jun Ni Bernhard P. Suter Jan Kubicek Barbara Maertens Thomas Schubert Camron Blackburn Shuguang Zhang |
| author_facet | Rui Qing Fei Tao Pranam Chatterjee Gaojie Yang Qiuyi Han Haeyoon Chung Jun Ni Bernhard P. Suter Jan Kubicek Barbara Maertens Thomas Schubert Camron Blackburn Shuguang Zhang |
| author_sort | Rui Qing |
| collection | DOAJ |
| description | Summary: It was posited that functionalities of GPCRs require full-length sequences that are negated by residue deletions. Here we report that significantly truncated nfCCR5QTY and nfCXCR4QTY still bind native ligands. Receptor-ligand interactions were discovered from yeast 2-hybrid screening and confirmed by mating selection. Two nfCCR5QTY (SZ218a, SZ190b) and two nfCXCR4QTY (SZ158a, SZ146a) were expressed in E. coli. Synthesized receptors exhibited α-helical structures and bound respective ligands with reduced affinities. SZ190b and SZ158a were reconverted into non-QTY forms and expressed in HEK293T cells. Reconverted receptors localized on cell membranes and functioned as negative regulators for ligand-induced signaling when co-expressed with full-length receptors. CCR5-SZ190b individually can perform signaling at a reduced level with higher ligand concentration. Our findings provide insight into essential structural components for CCR5 and CXCR4 functionality, while raising the possibility that non-full-length receptors may be resulted from alternative splicing and that pseudo-genes in genomes may be present and functional in living organisms. |
| first_indexed | 2024-12-14T05:41:18Z |
| format | Article |
| id | doaj.art-74a0381c3398455a98d7c52fd6f73be5 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-12-14T05:41:18Z |
| publishDate | 2020-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-74a0381c3398455a98d7c52fd6f73be52022-12-21T23:15:01ZengElsevieriScience2589-00422020-12-012312101670Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane ReceptorsRui Qing0Fei Tao1Pranam Chatterjee2Gaojie Yang3Qiuyi Han4Haeyoon Chung5Jun Ni6Bernhard P. Suter7Jan Kubicek8Barbara Maertens9Thomas Schubert10Camron Blackburn11Shuguang Zhang12Media Lab, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; Corresponding authorLaboratory of Food Microbial Technology, State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, ChinaMedia Lab, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; The Center for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USAMedia Lab, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USAMedia Lab, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USAMedia Lab, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USALaboratory of Food Microbial Technology, State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, ChinaNext Interactions, Inc., 2600 Hilltop Drive, Building B, C332, Richmond, CA 94806, USACube Biotech, GmbH, Creative Campus, Alfred-Nobel Strasse 10, 40789 Monheim, GermanyCube Biotech, GmbH, Creative Campus, Alfred-Nobel Strasse 10, 40789 Monheim, Germany2bind GmbH, Am BioPark 11, 93053 Regensburg, GermanyThe Center for Bits and Atoms, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USAMedia Lab, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; Corresponding authorSummary: It was posited that functionalities of GPCRs require full-length sequences that are negated by residue deletions. Here we report that significantly truncated nfCCR5QTY and nfCXCR4QTY still bind native ligands. Receptor-ligand interactions were discovered from yeast 2-hybrid screening and confirmed by mating selection. Two nfCCR5QTY (SZ218a, SZ190b) and two nfCXCR4QTY (SZ158a, SZ146a) were expressed in E. coli. Synthesized receptors exhibited α-helical structures and bound respective ligands with reduced affinities. SZ190b and SZ158a were reconverted into non-QTY forms and expressed in HEK293T cells. Reconverted receptors localized on cell membranes and functioned as negative regulators for ligand-induced signaling when co-expressed with full-length receptors. CCR5-SZ190b individually can perform signaling at a reduced level with higher ligand concentration. Our findings provide insight into essential structural components for CCR5 and CXCR4 functionality, while raising the possibility that non-full-length receptors may be resulted from alternative splicing and that pseudo-genes in genomes may be present and functional in living organisms.http://www.sciencedirect.com/science/article/pii/S2589004220308622Molecular BiologyCell BiologyStructural Biology |
| spellingShingle | Rui Qing Fei Tao Pranam Chatterjee Gaojie Yang Qiuyi Han Haeyoon Chung Jun Ni Bernhard P. Suter Jan Kubicek Barbara Maertens Thomas Schubert Camron Blackburn Shuguang Zhang Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors iScience Molecular Biology Cell Biology Structural Biology |
| title | Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors |
| title_full | Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors |
| title_fullStr | Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors |
| title_full_unstemmed | Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors |
| title_short | Non-full-length Water-Soluble CXCR4QTY and CCR5QTY Chemokine Receptors: Implication for Overlooked Truncated but Functional Membrane Receptors |
| title_sort | non full length water soluble cxcr4qty and ccr5qty chemokine receptors implication for overlooked truncated but functional membrane receptors |
| topic | Molecular Biology Cell Biology Structural Biology |
| url | http://www.sciencedirect.com/science/article/pii/S2589004220308622 |
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