title: Designed cyclic permutants of HIV-1 gp120: implications for envelope trimer structure and immunogen design.
abstract: Most HIV-1 broadly neutralizing antibodies are directed against the gp120 subunit of the env surface protein. Native env consists of a trimer of gp120-gp41 heterodimers, and in contrast to monomeric gp120, preferentially binds CD4 binding site (CD4bs)-directed neutralizing antibodies over non-neutralizing ones. Some cryo-electron tomography studies have suggested that the V1V2 loop regions of gp120 are located close to the trimer interface. We have therefore designed cyclically permuted variants of gp120 with and without the h-CMP and SUMO2a trimerization domains inserted into the V1V2 loop. h-CMP-V1cyc is one such variant in which residues 153 and 142 are the N- and C-terminal residues, respectively, of cyclically permuted gp120 and h-CMP is fused to the N-terminus. This molecule forms a trimer under native conditions and binds CD4 and the neutralizing CD4bs antibodies b12 with significantly higher affinity than wild-type gp120. It binds non-neutralizing CD4bs antibody F105 with lower affinity than gp120. A similar derivative, h-CMP-V1cyc1, bound the V1V2 loop-directed broadly neutralizing antibodies PG9 and PG16 with ∼20-fold higher affinity than wild-type JRCSF gp120. These cyclic permutants of gp120 are properly folded and are potential immunogens. The data also support env models in which the V1V2 loops are proximal to the trimer interface.
authors.affiliation: Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560 012, India.
authors.forename: Piyali
authors.initials: P
authors.lastname: Saha
authors.forename: Sanchari
authors.initials: S
authors.lastname: Bhattacharyya
authors.forename: Sannula
authors.initials: S
authors.lastname: Kesavardhana
authors.forename: Edward Roshan
authors.initials: ER
authors.lastname: Miranda
authors.forename: P Shaik Syed
authors.initials: PS
authors.lastname: Ali
authors.forename: Deepak
authors.initials: D
authors.lastname: Sharma
authors.forename: Raghavan
authors.initials: R
authors.lastname: Varadarajan
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000737:chemistry / Q000276:immunology; D001665:Binding Sites; D015704:CD4 Antigens / Q000737:chemistry / Q000276:immunology; D000939:Epitopes; D057809:HEK293 Cells; D015699:HIV Envelope Protein gp120 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000276:immunology; D055503:Protein Multimerization; D014162:Transfection; D014614:Vaccines, Synthetic / Q000737:chemistry* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D015704:CD4 Antigens; D000939:Epitopes; D015699:HIV Envelope Protein gp120; D010456:Peptides, Cyclic; D014614:Vaccines, Synthetic