5JO5

Crystal structure of 10E8 gHV-gLV antigen-binding fragment.

Summary for 5JO5

• Entry DOI: 10.2210/pdb5jo5/pdb
• Descriptor: 10E8 gHV, 10E8 gLV (3 entities in total)
• Functional Keywords: mper, hiv-1, antibody development, neutralizing, immune system
• Biological source: Homo sapiens; More
• Total number of polymer chains: 8
• Total formula weight: 189674.81

Authors

Joyce, M.G.,Mascola, J.R.,Kwong, P.D. (deposition date: 2016-05-02, release date: 2016-07-13, Last modification date: 2023-09-27)

Primary citation

Soto, C.,Ofek, G.,Joyce, M.G.,Zhang, B.,McKee, K.,Longo, N.S.,Yang, Y.,Huang, J.,Parks, R.,Eudailey, J.,Lloyd, K.E.,Alam, S.M.,Haynes, B.F.,Mullikin, J.C.,Connors, M.,Mascola, J.R.,Shapiro, L.,Kwong, P.D.
Developmental Pathway of the MPER-Directed HIV-1-Neutralizing Antibody 10E8.
Plos One, 11:e0157409-e0157409, 2016

PubMed Abstract: Antibody 10E8 targets the membrane-proximal external region (MPER) of HIV-1 gp41, neutralizes >97% of HIV-1 isolates, and lacks the auto-reactivity often associated with MPER-directed antibodies. The developmental pathway of 10E8 might therefore serve as a promising template for vaccine design, but samples from time-of-infection-often used to infer the B cell record-are unavailable. In this study, we used crystallography, next-generation sequencing (NGS), and functional assessments to infer the 10E8 developmental pathway from a single time point. Mutational analysis indicated somatic hypermutation of the 2nd-heavy chain-complementarity determining region (CDR H2) to be critical for neutralization, and structures of 10E8 variants with V-gene regions reverted to genomic origin for heavy-and-light chains or heavy chain-only showed structural differences >2 Å relative to mature 10E8 in the CDR H2 and H3. To understand these developmental changes, we used bioinformatic sieving, maximum likelihood, and parsimony analyses of immunoglobulin transcripts to identify 10E8-lineage members, to infer the 10E8-unmutated common ancestor (UCA), and to calculate 10E8-developmental intermediates. We were assisted in this analysis by the preservation of a critical D-gene segment, which was unmutated in most 10E8-lineage sequences. UCA and early intermediates weakly bound a 26-residue-MPER peptide, whereas HIV-1 neutralization and epitope recognition in liposomes were only observed with late intermediates. Antibody 10E8 thus develops from a UCA with weak MPER affinity and substantial differences in CDR H2 and H3 from the mature 10E8; only after extensive somatic hypermutation do 10E8-lineage members gain recognition in the context of membrane and HIV-1 neutralization.

PubMed: 27299673
DOI: 10.1371/journal.pone.0157409

Experimental method

X-RAY DIFFRACTION (1.7 Å)