Effect of surface electrostatic interactions on the stability and folding of formate dehydrogenase from Candida methylica |
| |
| Institution: | 1. Istanbul Technical University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul, Turkey;2. Istanbul Technical University, Molecular Biology-Biotechnology & Genetics Research Center, 34469 Istanbul, Turkey;3. Y?ld?z Technical University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul, Turkey;4. University of Bristol, Department of Biochemistry, Bristol BS8 1TD, Avon, United Kingdom;1. State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;2. University of Chinese Academy of Sciences, Beijing 100049, China;1. Institute for Vascular Signalling, Centre for Molecular Medicine, Goethe University, Frankfurt am Main, D-60596, Germany;2. German Center of Cardiovascular Research (DZHK) Partner site Rhein-Main, Frankfurt am Main, Germany;3. Division of Intramural Research, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, USA;4. Max-Delbrück Center for Molecular Medicine, Robert-Rössle-Str 10, 13092 Berlin, Germany;1. Department of Neurology, Boston Children''s Hospital, Harvard Medical School, Boston, MA, USA;2. Department of Neurology, New York University Comprehensive Epilepsy Center, New York University Langone Medical Center, New York University School of Medicine, New York, NY, USA;3. Department of Child Neurology, Hospital Sant Joan de Déu, Universidad de Barcelona, Barcelona, Spain;4. Epilepsy Center, Neurological Institute, Cleveland Clinic, Cleveland, OH, USA;5. Department of Neurology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA;1. Biochemistry Research Division, Faculty of Mathematics and Natural Sciences, Bandung Institute of Technology, Jalan Ganesa No 10, Bandung 40132, Indonesia;2. Center for Life Sciences, Bandung Institute of Technology, Jalan Ganesa No. 10, Bandung 40132, Indonesia;3. Laboratory of Biophysical Chemistry, University of Groningen, Nijenborgh 7, 9747 AG Groningen, The Netherlands;4. Biochemistry Laboratory, Department of Chemistry, Faculty of Mathematics and Natural Sciences, Padjadjaran University, Jalan Singaperbangsa No. 2, Bandung 40133, Indonesia;5. Mass Spectrometry Core Facility, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands |
| |
| Abstract: | NAD+-dependent formate dehydrogenase (FDH-EC 1.2.1.2) is an important enzyme to regenerate valuable NADH required by NAD+-dependent oxidoreductases in enzyme catalysis. The limitation in the thermostability of FDH enzyme is a crucial problem for development of biotechnological and industrial processes, despite of its advantages. In this study, to investigate the contribution of surface electrostatic interaction to the thermostability of FDH from Candida methylica (cmFDH) N187E, H13E, Q105R, N300E, N147R N300E/N147R, N187E/Q105R, N187E/N147R,Y160R, Y302R, Y160E and Y302E mutants were designed using a homology model of cmFDH based on Candida boidinii (cb) by considering electrostatic interactions on the protein surface. The effects of site-specific engineering on the stability of this molecule was analyzed according to minimal model of folding and assembly reaction and deduced equilibrium properties of the native system with respect to its thermal and denaturant sensitivities. It was observed that mutations did not change the unfolding pattern of native cmFDH and increased numbers of electrostatic interactions can cause either stabilizing or destabilizing effect on the thermostability of this protein. The thermodynamic and kinetic results suggested that except relatively improved mutants, three out of the nine single mutations increased the melting temperature of cmFDH enzyme. |
| |
| Keywords: | Electrostatic interaction Formate dehydrogenase Protein folding Enzyme stability |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
