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Defining specific residue‐to‐residue interactions between the gp120 bridging sheet and the N‐terminal segment of CCR 5: applications of transferred NOE NMR
Author(s) -
Srivastava Gautam,
Moseri Adi,
Kessler Naama,
Arshava Boris,
Naider Fred,
Anglister Jacob
Publication year - 2018
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.14673
Subject(s) - residue (chemistry) , chemistry , peptide , stereochemistry , nuclear magnetic resonance spectroscopy , valine , docking (animal) , isoleucine , tyrosine , biochemistry , protein structure , biophysics , amino acid , leucine , biology , medicine , nursing
Infection by HIV ‐1 requires protein–protein interactions involving gp120, CD 4 and CCR 5. We have previously demonstrated that the transferred nuclear Overhauser effect ( TRNOE ), in combination with asymmetric deuteration of a protein and a peptide ligand can be used to detect intermolecular interactions in large protein complexes with molecular weights up to ~ 100 kD a. Here, using this approach, we reveal interactions between tyrosine residues of a 27‐residue peptide corresponding to the N‐terminal segment of the CCR 5 chemokine receptor, and a dimeric extended core YU 2 gp120 envelope protein of HIV ‐1 complexed with a CD 4‐mimic miniprotein. The TRNOE crosspeaks in the ternary complex were assigned to the specific Tyr protons in the CCR 5 peptide and to methyl protons of isoleucine, leucine and/or valine residues of gp120. Site directed mutagenesis combined with selective deuteration and TRNOE resulted in the first discernment by a biophysical method of specific pairwise interactions between gp120 residues in the bridging sheet of gp120 and the N‐terminus of CCR 5.