Premium
Measurement of the Binding of Adenosylcobalamin to Glutamate Mutase by Fluorescence Spectroscopy
Author(s) -
Lin HengJu,
Hsu HuiJu,
Duann YehFang,
Chen HaoPing
Publication year - 2012
Publication title -
journal of the chinese chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.329
H-Index - 45
eISSN - 2192-6549
pISSN - 0009-4536
DOI - 10.1002/jccs.201100586
Subject(s) - adenosylcobalamin , chemistry , mutase , fluorescence , fluorescence spectroscopy , spectroscopy , cofactor , photochemistry , biochemistry , enzyme , physics , quantum mechanics
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 K d 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 B 12 to glutamate mutase using fluorescence spectroscopy.