On the effects of replacing the carboxylate-binding arginine-171 by hydrophobic tyrosine or tryptophan residues in the L-lactate dehydrogenase from Bacillus stearothermophilus

Biochemistry ◽  
1989 ◽  
Vol 28 (16) ◽  
pp. 6605-6610 ◽  
Author(s):  
Marcel A. Luyten ◽  
Marvin Gold ◽  
James D. Friesen ◽  
J. Bryan Jones
Keyword(s):  
Lactate Dehydrogenase ◽  
Bacillus Stearothermophilus ◽  
Tryptophan Residues

Site-directed mutagenesis of Bacillus stearothermophilus lactate dehydrogenase

1987 ◽  
Vol 15 (1) ◽  
pp. 152-153 ◽  
Author(s):  
ANTHONY R. CLARKE ◽  
DALE B. WIGLEY ◽  
DAVID A. BARSTOW ◽  
WILLIAM N. CHIA ◽  
ADAM D. B. WALDMAN ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Bacillus Stearothermophilus ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

Design and synthesis of new enzymes based on the lactate dehydrogenase framework

1991 ◽  
Vol 332 (1263) ◽  
pp. 177-184 ◽  
Keyword(s):  
Crystal Structure ◽  
Lactate Dehydrogenase ◽  
Malate Dehydrogenase ◽  
Functional Groups ◽  
Maximum Rate ◽  
Chemical Probes ◽  
Protein Surface ◽  
Design And Synthesis ◽  
Tryptophan Residues ◽  
Lactate Dehydrogenases

Analysis of the mechanism and structure of lactate dehydrogenases is summarized in a map of the catalytic pathway. Chemical probes, single tryptophan residues inserted at specific sites and a crystal structure reveal slow movements of the protein framework that discriminate between closely related small substrates. Only small and correctly charged substrates allow the protein to engulf the substrate in an internal vacuole that is isolated from solvent protons, in which water is frozen and hydride transfer is rapid. The closed vacuole is very sensitive to the size and charge of the substrate and provides discrimination between small substrates that otherwise have too few functional groups to be distinguished at a solvated protein surface. This model was tested against its ability to successfully predict the design and synthesis of new enzymes such as L-hydroxyisocaproate dehydrogenase and fully active malate dehydrogenase. Solvent friction limits the rate of forming the vacuole and thus the maximum rate of catalysis.

On the effect on specificity of Thr246→Gly mutation in L-lactate dehydrogenase of Bacillus stearothermophilus

1989 ◽  
Vol 161 (1) ◽  
pp. 59-63 ◽  
Author(s):  
Daniel Bur ◽  
Tony Clarke ◽  
James D. Friesen ◽  
Marvin Gold ◽  
Keith W. Hart ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Bacillus Stearothermophilus
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