期刊
FASEB JOURNAL
卷 29, 期 1, 页码 70-80出版社
FEDERATION AMER SOC EXP BIOL
DOI: 10.1096/fj.14-252262
关键词
crystal structure; entry; HIV; single-chain variable fragment
资金
- Grants-in-Aid for Scientific Research [24591484] Funding Source: KAKEN
- Canadian Institutes of Health Research Funding Source: Medline
- Howard Hughes Medical Institute Funding Source: Medline
- NIAID NIH HHS [AI094715, AI100890, R01 AI076119, R21 AI112417, AI076119, AI099284, AI112417, R21 AI094715, R01 AI100890, R37 AI076119, R01 AI099284] Funding Source: Medline
- NIGMS NIH HHS [P50 GM103368, GM103368] Funding Source: Medline
Humanized monoclonal antibody KD-247 targets the Gly(312)-Pro(313)-Gly(314)-Arg(315) arch of the third hypervariable (V3) loop of the HIV-1 surface glycoprotein. It potently neutralizes many HIV-1 clade B isolates, but not of other clades. To understand the molecular basis of this specificity, we solved a high-resolution (1.55 angstrom) crystal structure of the KD-247 antigen binding fragment and examined the potential interactions with various V3 loop targets. Unlike most antibodies, KD-247 appears to interact with its target primarily through light chain residues. Several of these interactions involve Arg(315) of the V3 loop. To evaluate the role of light chain residues in the recognition of the V3 loop, we generated 20 variants of KD-247 single-chain variable fragments with mutations in the antigen-binding site. Purified proteins were assessed for V3 loop binding using Alpha-Screen technology and for HIV-1 neutralization. Our data revealed that recognition of the clade-specificity defining residue Arg(315) of the V3 loop is based on a network of interactions that involve Tyr(L32), Tyr(L92), and Asn(L27d) that directly interact with Arg(315), thus elucidating the molecular interactions of KD-247 with its V3 loop target.
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