HIV-1 PROTEASE Mike Krzyskowski. Intro, Background HIV-1 – Human Immunodeficiency Virus 1 Highly...
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Transcript of HIV-1 PROTEASE Mike Krzyskowski. Intro, Background HIV-1 – Human Immunodeficiency Virus 1 Highly...
![Page 1: HIV-1 PROTEASE Mike Krzyskowski. Intro, Background HIV-1 – Human Immunodeficiency Virus 1 Highly virulent, highly contagious Left untreated progresses.](https://reader035.fdocuments.in/reader035/viewer/2022070411/56649f475503460f94c69625/html5/thumbnails/1.jpg)
HIV-1 PROTEASEMike Krzyskowski
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Intro, Background• HIV-1 – Human Immunodeficiency Virus 1
• Highly virulent, highly contagious• Left untreated progresses to AIDS
• About 1.6 million deaths worldwide in 2012 (WHO)
• Integral protein in HIV-1 lifecycle
• “chops” polyprotein into
appropriate size
Image: HIV-1 Protease (pdb 3FX5)
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Sequence Alignment
• HIV-1 protease, colored by chain
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Structure
• Red: Flap region• Orange: Active site triad (Asp-Thr-Gly)• Purple: Helices• Black: AA 93, mutation I93L implicated in drug resistance
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Structure: Active Site
KNI-272 Inhibitor
Asp25 (protonated)
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Mechanism• Hydrolysis
• H2O molecule held in active site (purple)
Asp25Asp125
KNI-272
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Mechanism• Only 1 Asp is protonated
• Asp farther from H2O
1. Negative Asp activates H2O
2. Attacks Carbonyl• Generation of oxyanion tetrahedral intermediate
3. Protonation of scissile N atom• Breakdown to hydrolysis products
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Drug Resistance: Not so simple• ANAM-11
• “Wild type”
• A-1• Active site
mutation
• NAM-10• Non-active site
mutations
Image source: Muzzamil et, al. (2003)
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Drug Resistance
• Substrates: synthetic proteins that mimic natural cleavage sites
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References• Adachi, M., Ohhara, T., Kurihara, K., Tamada, T., Honjo, E., Okazaki, N., Arai, S., Shoyama, Y., Kimura, K., Matsumura, H., Sugiyama, S., Adachi,
H., Takano, K., Mori, Y., Hidaka, K., Kimura, T., Hayashi, Y., Kiso, Y., Kuroki, R. (2009) Structure of HIV-1 Protease in Complex with Potent Inhibitor KNI-272 Determined by High Resolution X-ray Crystallography Proc.Natl.Acad.Sci.USA
• Brik, A and Wong, C. (2003) HIV-1 Protease: Mechanism and Drug Discovery. Org. Bio.Chem. 1: 5-14
• Hidaka, K., Kimura, T., Abdel-Rahman, H.M., Nguyen, J.T., McDaniel, K.F., Kohlbrenner, W.E., Molla, A., Adachi, M., Tamada, T., Kuroki, R., Katsuki, N., Tanaka, Y., Matsumoto, H., Wang, J., Hayashi, Y., Kempf, D.J., Kiso, Y. (2009) Crystal Structure of HIV-1 Protease Complexed with KNI-1689. J.Med.Chem. 52: 7604-7617
• Jaskólski, M, Tomasselli, G Sawyer, T K Staples, D G Heinrikson, R L Schneider, J Kent, S B, Wlodawer, A Structure at 2.5-A resolution of chemically synthesized human immunodeficiency virus type 1 protease complexed with a hydroxyethylene-based inhibitor. (1991) 30: 1600-9
• Kohl, N E Emini, E a Schleif, W a Davis, L J Heimbach, J C Dixon, R a Scolnick, E M Sigal, I S Active human immunodeficiency virus protease is required for viral infectivity. (1988) Proc. Nat. Acad. Sci. 85: 4686-90
• Kovalevsky, A.Y., Liu, F., Leshchenko, S., Ghosh, A.K., Louis, J.M., Harrison, R.W., Weber, I.T. (2006) Ultra-high resolution X-ray crystal structure of HIV-1 protease V32I mutant with TMC114 (darunavir) inhibitor J.Mol.Biol. 363: 161-173
• Muzammil, S Ross, P and Freire, E. A Major Role for a Set of Non-Active Site Mutations in the Development of HIV-1 Protease Drug Resistance (2003) Biochemistry 42:631-638
• Servais, J., Lambert, C., Fontaine, E., Plesse, J., Robert, I., Arendt, V. I. C., … Schmit, J. (2001). Variant human immunodeficiency virus type 1 proteases and response to combination therapy including a protease inhibitor. Antimicrobial Agents …, 45(3), 893–900.
• Strisovsky K, Tessmer U, Langner J, Konvalinka J, Kräusslich HG.. Systematic mutational analysis of the active-site threonine of HIV-1 proteinase: rethinking the "fireman's grip" hypothesis. (2000) Pro. Sci 9(9):1631-41.
• K. Suguna, Richard R. Bott†, Eduardo A. Padlan, E. Subramanian‡,, Steven Sheriff,, Gerson H. Cohen, Davies David (1987) Structure and refinement at 1.8 Å resolution of the aspartic proteinase from Rhizopus chinensis J. Mol. Bio. 196(4):877-900
• http://www.who.int/gho/hiv/en/