title: The expression and purification of WSSV134 from white spot syndrome virus and its inhibitory effect on caspase activity from Penaeus monodon.
abstract: WSSV134 or VP36A protein of white spot syndrome virus was previously reported to be able to reduce apoptosis in Sf-9 cells transfected with caspase of Penaeus monodon (PmCasp). The protein was therefore believed to have a role in supporting the survival of WSSV inside the host cells during infection. However, the anti-apoptosis activity of WSSV134 involved in the inhibition of PmCasp is still unclear. In this study, we produced a recombinant WSSV134 (rWSSV134) and tested for its ability to inhibit PmCasp in vitro. The results from a caspase inhibition assay revealed that rWSSV134 could inhibit PmCasp in a dose-dependent manner. Since WSSV134 was predicted to contain three potential caspase binding sites, corresponding to the D54, D104 and D259, we then employed site-directed mutagenesis to investigate the involvement of these sites in PmCasp inhibition. D54A and D259A mutants could still inhibit PmCasp while D104A mutant lacks this activity. Our results confirmed that the WSSV134 is an inhibitor for PmCasp and that residue D104 is important for PmCasp inhibition.
authors.affiliation: Department of Microbiology, Faculty of Science, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand.
authors.forename: Thanunthon
authors.initials: T
authors.lastname: Bowornsakulwong
authors.affiliation: Center of Excellence for Shrimp Molecular Biology and Biotechnology (Centex Shrimp), Faculty of Science, Mahidol University, Bangkok 10400, Thailand; National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani 12120, Thailand.
authors.forename: Walaiporn
authors.initials: W
authors.lastname: Charoensapsri
authors.affiliation: Department of Microbiology, Faculty of Science, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand.
authors.forename: Triwit
authors.initials: T
authors.lastname: Rattanarojpong
authors.affiliation: Department of Microbiology, Faculty of Science, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand. Electronic address: pongsak.khu@kmutt.ac.th.
authors.forename: Pongsak
authors.initials: P
authors.lastname: Khunrae
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D061945:Caspase Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D020169:Caspases / Q000737:chemistry*; D033561:Penaeidae / Q000201:enzymology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D061987:Sf9 Cells; D018411:Spodoptera; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D046848:White spot syndrome virus 1 / Q000737:chemistry / Q000235:genetics*
chemical_list: D060829:Arthropod Proteins; D061945:Caspase Inhibitors; D011994:Recombinant Proteins; D014764:Viral Proteins; D020169:Caspases
keywords: Apoptosis; Caspase; WSSV134; White spot syndrome virus