title: Large changes of transition-state structure during experimental evolution of an enzyme.
abstract: The question of whether, during the evolution of an enzyme, the transition state of the catalysed reaction is largely unchanged, or whether transition state and protein change together, was examined using the egb beta-galactosidases of Escherichia coli. Charge development at the first chemical state was assumed [Konstantinidis and Sinnott (1991) Biochem. J. 279, 587-593] to be proportional to delta delta G++, the ratio of second-order rate constants for the hydrolysis of beta-D-galactopyranosyl fluoride and 1-fluoro-D-galactopyranosyl fluoride, expressed as a free-energy difference. delta delta G++ (kJ.mol-1) falls from 10.4 for wild-type enzyme to 6.8 and 7.2 as a consequence of two different single amino-acid changes (which arise from single evolutionary events), to 6.3 as a consequence of the two amino-acid changes together, and then increases slightly to 7.3 as a consequence of a third single evolutionary change involving three further amino-acid changes.
authors: Department of Chemistry, University of Illinois, Chicago 60607-7061.
authors: K
authors: K
authors: Srinivasan
authors: A
authors: A
authors: Konstantinidis
authors: M L
authors: ML
authors: Sinnott
authors: B G
authors: BG
authors: Hall
mesh_terms: D000595:Amino Acid Sequence; D005075:Biological Evolution; D002384:Catalysis; D004926:Escherichia coli / Q000201:enzymology*; D006868:Hydrolysis; D007700:Kinetics; D013816:Thermodynamics; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D001616:beta-Galactosidase