title: Engineering Metamorphic Chemokine Lymphotactin/XCL1 into the GAG-Binding, HIV-Inhibitory Dimer Conformation.
abstract: Unlike other chemokines, XCL1 undergoes a distinct metamorphic interconversion between a canonical monomeric chemokine fold and a unique β-sandwich dimer. The monomeric conformation binds and activates the receptor XCR1, whereas the dimer binds extracellular matrix glycosaminoglycans and has been associated with anti-human immunodeficiency virus (HIV) activity. Functional studies of WT-XCL1 are complex, as both conformations are populated in solution. To overcome this limitation, we engineered a stabilized dimeric variant of XCL1 designated CC5. This variant features a new disulfide bond (A36C-A49C) that prevents structural interconversion by locking the chemokine into the β-sandwich dimeric conformation, as demonstrated by NMR structural analysis and hydrogen/deuterium exchange experiments. Functional studies analyzing glycosaminoglycan binding demonstrate that CC5 binds with high affinity to heparin. In addition, CC5 exhibits potent inhibition of HIV-1 activity in primary peripheral blood mononuclear cells (PBMCs), demonstrating the importance of the dimer in blocking viral infection. Conformational variants like CC5 are valuable tools for elucidating the biological relevance of the XCL1 native-state interconversion and will assist in future antiviral and functional studies.
authors: Department of Biochemistry, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors: Jamie C
authors: JC
authors: Fox
authors: Department of Biochemistry, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors: Robert C
authors: RC
authors: Tyler
authors: Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors: Christina
authors: C
authors: Guzzo
authors: Department of Biochemistry, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors: Robbyn L
authors: RL
authors: Tuinstra
authors: Department of Biochemistry, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors: Francis C
authors: FC
authors: Peterson
authors: Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health , Bethesda, Maryland 20892, United States.
authors: Paolo
authors: P
authors: Lusso
authors: Department of Biochemistry, Medical College of Wisconsin , Milwaukee, Wisconsin 53226, United States.
authors: Brian F
authors: BF
authors: Volkman
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use; D019744:Chemokines, C / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D019281:Dimerization; D014644:Genetic Variation; D006025:Glycosaminoglycans / Q000737:chemistry*; D015658:HIV Infections / Q000188:drug therapy; D015497:HIV-1 / Q000187:drug effects; D006493:Heparin / Q000737:chemistry; D006801:Humans; D008222:Lymphokines / Q000737:chemistry*; D011485:Protein Binding; D015202:Protein Engineering; D012795:Sialoglycoproteins / Q000737:chemistry*; D013329:Structure-Activity Relationship
chemical_list: D019380:Anti-HIV Agents; D019744:Chemokines, C; D006025:Glycosaminoglycans; D008222:Lymphokines; D012795:Sialoglycoproteins; C092390:XCL1 protein, human; C018997:lymphotactin; D006493:Heparin