Truncation of the gp41 construct just past its transmembrane domain, leaving residues 1-190, results in a well-structured protein, soluble in dodecylmaltoside while adopting its natural homo-trimeric form. The protein is found to be quite stable, even at temperatures of 50C, and samples are found to be suitable for NMR spectroscopy. Preliminary analysis shows that even thought the ecto-domain resonances are severely broadened, characteristic of slow molecular tumbling, the fusion domains of the trimer yield good NMR spectral characteristics. We find that these fusion domains exhibit chemical shifts that fall very close to those of the isolated domain, previously studied in SDS micelles and found to be uniformly alpha-helical. The fusion domains exhibit effective correlation times that are much shorter than for the remainder of the large trimeric complex, indicating that the intact fusion domain helices are highly mobile within the detergent micelle, while retaining their helical conformation. This excludes the previously hypothesized interaction with the gp41 transmembrane domain.
| Chiliveri, Sai Chaitanya; Louis, John M; Ghirlando, Rodolfo et al. (2018) Tilted, Uninterrupted, Monomeric HIV-1 gp41 Transmembrane Helix from Residual Dipolar Couplings. J Am Chem Soc 140:34-37 |
| Roche, Julien; Louis, John M; Aniana, Annie et al. (2015) Complete dissociation of the HIV-1 gp41 ectodomain and membrane proximal regions upon phospholipid binding. J Biomol NMR 61:235-48 |
| Roche, Julien; Louis, John M; Grishaev, Alexander et al. (2014) Dissociation of the trimeric gp41 ectodomain at the lipid-water interface suggests an active role in HIV-1 Env-mediated membrane fusion. Proc Natl Acad Sci U S A 111:3425-30 |
| Lakomek, Nils-Alexander; Kaufman, Joshua D; Stahl, Stephen J et al. (2013) Internal dynamics of the homotrimeric HIV-1 viral coat protein gp41 on multiple time scales. Angew Chem Int Ed Engl 52:3911-5 |
| Lakomek, Nils-Alexander; Ying, Jinfa; Bax, Ad (2012) Measurement of ยน?N relaxation rates in perdeuterated proteins by TROSY-based methods. J Biomol NMR 53:209-21 |
