Measurement of the Binding of Adenosylcobalamin to Glutamate Mutase by Fluorescence Spectroscopy |
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| Authors: | Heng‐Ju Lin Hui‐Ju Hsu Yeh‐Fang Duann Hao‐Ping Chen |
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| Affiliation: | 1. Institute of Biotechnology, National Taipei University of Technology, Taipei 106, Taiwan;2. Department of Biochemistry, School of Medicine, Tzu Chi University, Hualian 970, Taiwan |
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| Abstract: | Fluorescence spectroscopy is a fast, highly sensitive technique for investigating protein‐ligand interactions. Intrinsic protein fluorescence is usually occurred by exciting the proteins with 280‐295 nm ultraviolet light, and the light emission is observed approximately between 330‐350 nm. No emission light between 330‐350 nm can be observed when adenosylcobalamin (AdoCbl) is excited at 282 nm. The binding of AdoCbl to glutamate mutase was therefore investigated by fluorescence spectroscopy in this study. Our results show that direct measurement for determining the Kd of AdoCbl by fluorescence spectroscopy leads to significant errors. Here we report the source of error and a corrected method for measuring the binding of coenzyme B12 to glutamate mutase using fluorescence spectroscopy. |
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| Keywords: | Adenosylcobalamin Fluorescence spectroscopy Glutamate mutase |
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