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Ketterman, Albert J.. Active site residues of Dengue NS5 Guanylyltransferase. (). King Mongkut's University of Technology North Bangkok. Central Library. : , 2021.
Active site residues of Dengue NS5 Guanylyltransferase
Name: Isara Nachampa
Organization :
Mahidol University. Institute of Molecular Biosciences
Name: Chonticha Saisawang
Organization :
Mahidol University. Institute of Molecular Biosciences
Name: Ketterman, Albert J.
Organization :
Mahidol University. Institute of Molecular Biosciences
ThaSH:
Guanylyltransferase
ThaSH:
Dengue viruses
-- Prevention and control
ThaSH:
Epidemiology
Abstract:
This study aims to investigate and identify the critical amino acid residues
which can affect the structure, catalysis, or binding affinity of guanylyltransferase, an
important enzyme role in the viral RNA capping process. PCR site-directed
mutagenesis was performed to engineer alanine substitutions for S150 and R212
positions in the capping domain. Wild-type and mutated proteins were purified by
Maltose Binding Protein (MBP) column and HisTrap column. All purified capping
enzymes; wild-type, S150A, and R212A, were used to perform GTase assay using GTP
labeled with the fluorescent dye as a substrate. Michaelis-Menten plots were generated
for capping enzyme activity versus substrate concentration for the enzymes. All data
plots were analyzed using Prism5, yielding Vmax of mutants which are S150A
(0.0450±0.0158) and R212A (0.0377±0.0166) which are not significantly different
from the wild-type (0.0607±0.0044) but might have contribute in catalysis. Likewise,
KM of S150A (0.2691±0.1283 μM) and R212A (0.2384±0.1441 μM) are not significant
lower than its wild-type (0.3441±0.1100 μM). This data suggests those residues (S150
and R212) might have small contributions to binding affinity.
King Mongkut's University of Technology North Bangkok. Central Library
Address:
BANGKOK
Email:
library@kmutnb.ac.th
Created:
2021
Modified:
2023-06-09
Issued:
2023-06-09
BibliograpyCitation :
In Thai Society for Biotechnology. Thai Society for Biotechnology and International Conference Online (TSB Online 2021) (pp.599-603). Bangkok : Thai Society for Biotechnology
eng
©copyrights
RightsAccess:
ใช้เวลา
0.014893 วินาที
Isara Nachampa
| Title | Contributor | Type |
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Chonticha Saisawang
| Title | Contributor | Type |
|---|
Ketterman, Albert J.
| Title | Contributor | Type |
|---|