Computational design of a substrate specificity mutant of a protein.
The wild-type trp repressor of E. coli bound 5-methoxytryptophan, a Trp analogue, less tightly than Trp. A mutant repressor (Val58-->Ala) that should bind 5-methoxytryptophan preferentially to Trp was computationally designed by free-energy calculations accompanied by free-energy decomposition. The designed mutant was demonstrated by experiments to bind 5-methoxytryptophan more tightly than Trp, consistent with the computational prediction. This success indicates the usefulness of free energy decomposition in protein design.
Honda
N
N
Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan.
Komeiji
Y
Y
Uebayasi
M
M
Yamato
I
I
D001426:Bacterial Proteins; D008956:Models, Chemical; D016296:Mutagenesis; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013816:Thermodynamics; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000378:metabolism; D029968:Escherichia coli Proteins
D001426:Bacterial Proteins; D012097:Repressor Proteins; D014364:Tryptophan; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins; C027986:5-methoxytryptophan