An evolutionary approach to the design of glutathione-linked enzymes.
Studies of protein structure provide information about principles of protein design that have come into play in natural evolution. This information can be exploited in the redesign of enzymes for novel functions. The glutathione-binding domain of glutathione transferases has similarities with structures in other glutathione-linked proteins, such as glutathione peroxidases and thioredoxin (glutaredoxin), suggesting divergent evolution from a common ancestral protein fold. In contrast, the binding site for glutathione in human glyoxalase I is located at the interface between the two identical subunits of the protein. Comparison with the homologous, but monomeric, yeast glyoxalase I suggests that new domains have originated through gene duplications, and that the oligomeric structure of the mammalian glyoxalase I has arisen by 'domain swapping'. Recombinant DNA techniques are being used for the redesign of glutathione-linked proteins in attempts to create binding proteins with novel functions and catalysts with tailored specificities. Enzymes with desired properties are selected from libraries of variant structures by use of phage display and functional assays.
Mannervik
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B
Department of Biochemistry, Uppsala University, Sweden. bengt.mannervik@bmc.uu.se
Cameron
A D
AD
Fernandez
E
E
Gustafsson
A
A
Hansson
L O
LO
Jemth
P
P
Jiang
F
F
Jones
T A
TA
Larsson
A K
AK
Nilsson
L O
LO
Olin
B
B
Pettersson
P L
PL
Ridderström
M
M
Stenberg
G
G
Widersten
M
M
D001665:Binding Sites / Q000235:genetics; D019020:Directed Molecular Evolution; D015195:Drug Design; D004798:Enzymes / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism*; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007791:Lactoylglutathione Lyase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D015202:Protein Engineering; D017510:Protein Folding
D004798:Enzymes; D005982:Glutathione Transferase; D007791:Lactoylglutathione Lyase; D005978:Glutathione