Time-dependent XAS studies of trapped enzyme-substrate complexes of alcohol dehydrogenase from Thermoanaerobacter brockii

• Published in: Journal of synchrotron radiation Vol. 8; no. Pt 2; p. 978
• Main Authors: Kleifeld, O, Frenkel, A, Sagi, I
• Format: Journal Article
• Language: English
• Published: United States 01-03-2001
• Subjects: Alcohol Dehydrogenase - chemistry; Alcohol Dehydrogenase - metabolism; Bacteria, Anaerobic - enzymology; Butanols - chemistry; Butanols - metabolism; Gram-Positive Asporogenous Rods, Irregular - enzymology; NADP - chemistry; NADP - metabolism; Spectrometry, X-Ray Emission - methods
• ISSN: 0909-0495
• DOI: 10.1107/S0909049500017684

The understanding of structure-function relationships in proteins has been significantly advanced with the advent of the biotechnological revolution. A goal yet to be realized for many metalloenzyme systems is to characterize the dynamic changes in structure that bridge the static endpoints provided by crystallography. We present here a series of edge and EXAFS spectra of the metalloenzyme alcohol dehydrogenase from Thermoanaerobacter brockii (TbADH) complexed with its substrate. The enzyme-substrate complexes were trapped by fast freezing at various times, following their enzyme activity. Our edge and EXAFS analyses both reveal the time-dependent changes in the structure of the active site of TbADH.