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Structural evidence for a ligand coordination switch in liver alcohol dehydrogenase

Meijers, Rob; Adolph, Hans-Werner; Dauter, Zbigniew; Wilson, Keith S.; Lamzin, Victor S. and Cedergren, Eila LU (2007) In Biochemistry 46(18). p.5446-5454
Abstract
The use of substrate analogues as inhibitors provides a way to understand and manipulate enzyme function. Here we report two 1 A resolution crystal structures of liver alcohol dehydrogenase in complex with NADH and two inhibitors: dimethyl sulfoxide and isobutyramide. Both structures present a dynamic state of inhibition. In the dimethyl sulfoxide complex structure, the inhibitor is caught in transition on its way to the active site using a flash-freezing protocol and a cadmium-substituted enzyme. One inhibitor molecule is partly located in the first and partly in the second coordination sphere of the active site metal. A hydroxide ion bound to the active site metal lies close to the pyridine ring of NADH, which is puckered in a twisted... (More)
The use of substrate analogues as inhibitors provides a way to understand and manipulate enzyme function. Here we report two 1 A resolution crystal structures of liver alcohol dehydrogenase in complex with NADH and two inhibitors: dimethyl sulfoxide and isobutyramide. Both structures present a dynamic state of inhibition. In the dimethyl sulfoxide complex structure, the inhibitor is caught in transition on its way to the active site using a flash-freezing protocol and a cadmium-substituted enzyme. One inhibitor molecule is partly located in the first and partly in the second coordination sphere of the active site metal. A hydroxide ion bound to the active site metal lies close to the pyridine ring of NADH, which is puckered in a twisted boat conformation. The cadmium ion is coordinated by both the hydroxide ion and the inhibitor molecule, providing structural evidence of a coordination switch at the active site metal ion. The structure of the isobutyramide complex reveals the partial formation of an adduct between the isobutyramide inhibitor and NADH. It provides evidence of the contribution of a shift from the keto to the enol tautomer during aldehyde reduction. The different positions of the inhibitors further refine the knowledge of the dynamics of the enzyme mechanism and explain how the crowded active site can facilitate the presence of a substrate and a metal-bound hydroxide ion. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemistry
volume
46
issue
18
pages
5446 - 5454
publisher
The American Chemical Society
external identifiers
  • wos:000246071700018
  • scopus:34248205214
ISSN
0006-2960
DOI
10.1021/bi6023594
language
English
LU publication?
yes
id
27aa1bd7-0bd5-4128-a995-fbb8feb3de41 (old id 663225)
date added to LUP
2007-12-13 14:19:24
date last changed
2017-10-01 03:43:17
@article{27aa1bd7-0bd5-4128-a995-fbb8feb3de41,
  abstract     = {The use of substrate analogues as inhibitors provides a way to understand and manipulate enzyme function. Here we report two 1 A resolution crystal structures of liver alcohol dehydrogenase in complex with NADH and two inhibitors: dimethyl sulfoxide and isobutyramide. Both structures present a dynamic state of inhibition. In the dimethyl sulfoxide complex structure, the inhibitor is caught in transition on its way to the active site using a flash-freezing protocol and a cadmium-substituted enzyme. One inhibitor molecule is partly located in the first and partly in the second coordination sphere of the active site metal. A hydroxide ion bound to the active site metal lies close to the pyridine ring of NADH, which is puckered in a twisted boat conformation. The cadmium ion is coordinated by both the hydroxide ion and the inhibitor molecule, providing structural evidence of a coordination switch at the active site metal ion. The structure of the isobutyramide complex reveals the partial formation of an adduct between the isobutyramide inhibitor and NADH. It provides evidence of the contribution of a shift from the keto to the enol tautomer during aldehyde reduction. The different positions of the inhibitors further refine the knowledge of the dynamics of the enzyme mechanism and explain how the crowded active site can facilitate the presence of a substrate and a metal-bound hydroxide ion.},
  author       = {Meijers, Rob and Adolph, Hans-Werner and Dauter, Zbigniew and Wilson, Keith S. and Lamzin, Victor S. and Cedergren, Eila},
  issn         = {0006-2960},
  language     = {eng},
  number       = {18},
  pages        = {5446--5454},
  publisher    = {The American Chemical Society},
  series       = {Biochemistry},
  title        = {Structural evidence for a ligand coordination switch in liver alcohol dehydrogenase},
  url          = {http://dx.doi.org/10.1021/bi6023594},
  volume       = {46},
  year         = {2007},
}