Chapter Twelve - Proton transfer and drug binding details revealed in neutron diffraction studies of wild-type and drug resistant HIV-1 protease [electronic resource]

• Ngôn ngữ: eng
• Ký hiệu phân loại: 614.5 Incidence of and public measures to prevent specific diseases and kinds of diseases
• Thông tin xuất bản: Oak Ridge, Tenn. : Oak Ridge, Tenn. : Oak Ridge National Laboratory ; Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy, 2020
• Mô tả vật lý: Size: p. 257-279 : , digital, PDF file.
• Bộ sưu tập: Metadata
• ID: 259901
• Chủ đề: 1. 60 applied life sciences; 2. Acquired immunodeficiency syndrome; 3. Clinical protease inhibitor; 4. Hiv-1 protease; 5. Human immunodeficiency virus type 1; 6. Hydrogen bond; 7. Neutron crystallography; 8. Proton transfer; 9. Protonation state; 10. Structure-assisted drug design

Tóm tắt

HIV-1 protease is an essential therapeutic target for the design and development of antiviral inhibitors to treat AIDS. We used room temperature neutron crystallography to accurately determine hydrogen atom positions in several protease complexes with clinical drugs, amprenavir and darunavir. Hydrogen bonding interactions were carefully mapped to provide an unprecedented picture of drug binding to the protease target. We demonstrate that hydrogen atom positions within the enzyme catalytic site can be altered by introducing drug resistant mutations and by protonating surface residues that trigger proton transfer reactions between the catalytic Asp residues and the hydroxyl group of darunavir. When protein perdeuteration is not feasible, we validate the use of initial H/D exchange with unfolded protein and partial deuteration in pure D2O with hydrogenous glycerol to maximize deuterium incorporation into the protein, with no detrimental effects on the growth of quality crystals suitable for neutron diffraction experiments.