Please use this identifier to cite or link to this item: http://hdl.handle.net/10603/4694
Title: X-ray crystallographic studies on HIV-1 protease
Researcher: Das, Amit
Guide(s): Hosur, M V
Keywords: crystal structure
HIV-1 protease
enzyme
catalysis
mechanism
drug resistance
substrate
RT-RH
TFR-PR
tetrahedral intermediate
Upload Date: 13-Sep-2012
University: Homi Bhabha National Institute
Completed Date: 09-03-2012
Abstract: The unique contribution of the candidate is structural mapping of the peptide bond cleavage reaction catalyzed by HIV-1 protease (HIV-1 PR). The candidate has obtained the first crystal structures of complexes between an active HIV-1 PR enzyme and natural type-1 and type-2 substrates. In the complexes with a type-2 substrate, the substrate is captured in the active site at three different stages of the cleavage reaction: when bound as a regular peptide, after in-situ modification into a tetrahedral reaction intermediate, and when cleaved into product peptides, which are still bound in the active site of the enzyme. He has discovered an inter-enzyme-substrate short ionic hydrogen bond (SIHB) in the tetrahedral intermediate complex, and an intra- enzyme low barrier hydrogen bond (LBHB) just after the substrate is cleaved into product peptides. In the complex with type-1 substrate, the substrate is cleaved and the C-terminal peptide bond about the proline residue displays cis conformation. These are very novel and atomic level descriptions of substrate recognition and processing by HIV-1 protease enzyme. Based on these structural inputs he has given a detailed mechanism of the peptide bond hydrolysis by HIV-1 PR. Through very high resolution structures, the candidate has found that in unliganded HIV-1 PR, the inter-aspartate hydrogen bond is not a LBHB, in contradiction to the latest mechanistic proposal. Further, the candidate has found novel arrangement of water molecules at the catalytic centre, only when the pH is non-optimal for enzyme activity. The rate reduction at non-optimal pH?s is suggested to be partly due to this water structure. The candidate has also determined the structures of the complexes of FDA approved drug, ritonavir with native as well as HIV-1 PR mutants that are resistant to ritonavir. These structures reveal that, in V82F mutant, loss of hydrophobic interactions contributes significantly toward development of resistance and the M36I mutant has a compensatory role in restoring viral fitness.
Pagination: 231p.
URI: http://hdl.handle.net/10603/4694
Appears in Departments:Department of Chemical Sciences

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01_title.pdfAttached File1.7 MBAdobe PDFView/Open
02_certificate.pdf414.55 kBAdobe PDFView/Open
03_declaration.pdf649.13 kBAdobe PDFView/Open
04_dedication.pdf359.25 kBAdobe PDFView/Open
05_acknowledgements.pdf318.72 kBAdobe PDFView/Open
06_contents.pdf359.33 kBAdobe PDFView/Open
07_synopsis.pdf579.95 kBAdobe PDFView/Open
08_list of figures.pdf298.95 kBAdobe PDFView/Open
09_list of tables.pdf194.02 kBAdobe PDFView/Open
10_chapter 1.pdf889.1 kBAdobe PDFView/Open
11_chapter 2.pdf1.17 MBAdobe PDFView/Open
12_chapter 3.pdf4.22 MBAdobe PDFView/Open
13_chapter 4.pdf2.09 MBAdobe PDFView/Open
14_chapter 5.pdf2.15 MBAdobe PDFView/Open
15_chapter 6.pdf3.4 MBAdobe PDFView/Open
16_references.pdf928.81 kBAdobe PDFView/Open
17_key words and abbreviations.pdf257.91 kBAdobe PDFView/Open
18_abstract.pdf81.96 kBAdobe PDFView/Open
19_list of publications.pdf314.77 kBAdobe PDFView/Open


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