Recombinant NS3 Seine Protease From Dengue Virus 2 As A Screen For Small Molecules

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dc.contributor.authorAb Aziz, Nurohaida
dc.date.accessioned2018-12-05T06:48:29Z
dc.date.available2018-12-05T06:48:29Z
dc.date.issued2011-08
dc.description.abstractDengue infection is re-emerging as a major global disease and is classified as a Category A priority pathogen. Dengue viruses are estimated to infect 50-100 million people annually and are considered to cause one of the most important arthropod-borne viral diseases in terms of human morbidity and mortality. Virus transmission occurs through the bite of the Aedes aegypti mosquito and half the world’s population is at risk for infection. There is presently no approved vaccine or antiviral drug that is effective against dengue viruses. The focus of this thesis is to combine the power of high performance computing with wet lab experiments for the recombinant NS3 serine protease from dengue virus type 2 as a screen for antiviral small molecules that can be used either to prevent or treat dengue virus infections. In collaboration with Dr. Irene Newhouse, Advance Studies for Genomics, Proteomics and Bioinformatics (ASGPB), University of Hawaii, molecular modelling and in silico screening of small molecule compound libraries from the National Cancer Institute (NCI) and ZINC databases that dock into the DENV2 NS2B-NS3 protease binding site was carried out. From the pool of best-fit candidates (53 potential small molecule inhibitors), the 4 high-scoring water-soluble, commercially available compounds were selected for in vitro assessment. The NS2B-NS3 serine protease gene from dengue virus serotype 2 was cloned and expressed in E. coli as a recombinant hexahistidine tagged protein. The NS2B-NS3 protease was purified using affinity and gel filtration chromatography. In vitro assay revealed protease activity toward a fluorogenic peptide substrate containing two basic amino acid residues. All 4 high-scoring water-soluble compounds were tested and exhibited in vitro inhibition activity on the recombinant NS2B-NS3 serine protease. Compound 4 was found to be most active inhibitor with 64% inhibition at 100 M concentration.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/7210
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectRecombinant NS3 seine proteaseen_US
dc.subjectfrom dengue virus 2en_US
dc.titleRecombinant NS3 Seine Protease From Dengue Virus 2 As A Screen For Small Moleculesen_US
dc.typeThesisen_US
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