FERSHT

Fersht A R 1985 Enzyme Struoture and Meohanism 2nd edn (New York Freeman)... 

AR Fersht. Structure and Mechanism in Protein Science A Guide to Enzyme Catalysis and Protein Folding. New York WFl Freeman, 1999. 

Figure 4.18 Side chains of the tyrosyl-tRNA synthetase that form hydrogen bonds to tyrosyl adenylate. Green residues are from p strand 2 and the following loop regions, yellow residues are from the loop after P strand S, and brown residues are from the a helix before P strand S. (Adapted from T. Wells and A. Fersht, Nature 316 656-657, 1985.)...

Recently Alan Fersht, Cambridge University, has developed a protein engineering procedure for such studies. The technique is based on investigation of the effects on the energetics of folding of single-site mutations in a protein of known structure. For example, if minimal mutations such as Ala to Gly in the solvent-exposed face of an a helix, destabilize both an intermediate state and the native state, as well as the transition state between them, it is likely that the helix is already fully formed in the intermediate state. If on the other hand the mutations destabilize the native state but do not affect the energy of the intermediate or transition states at all, it is likely that the helix is not formed until after the transition state. 

Fersht, A.R. Characterizing transition states in protein folding an essential step in the puzzle. Curr. Opin. Struct. Biol 5 79-84, 1994. 

Fersht, A. Enzyme Structure and Mechanism, 2nd ed. New York W.H. Freeman, 1984. 

Thomas, P.G., Russel, A.J., Fersht, A. Tailoring the pH dependence of enzyme catalysis using protein engineering. Nature 318 375-376, 1985. 

Fersht, A.R. The hydrogen bond in molecular recognition. Trends Biochem. Sci. 12 301-304, 1987. 

Fersht, A.R., et al. Hydrogen bonding and biological specificity analyzed by protein engineering. Nature 314 235-238, 1985. 

T. C. Bruice and S. I Benkovic, Bioorganic Mechanisms, Vol. 1, W. A. Benjamin, New brk, 1966, pp. 1-258 W. P. Jencks, Catalysis in Chemistry and Enzymology, McGraw-Hill, New York, 1969 M. L. Bender, Mechanisms of Homogeneous Catalysis from Protons to Proteins, Wiley-Interscience, New York, 1971 C. Walsh, Enzymatic Reaction Mechanisms, W. H. Freeman, San Francisco, 1979 A. Fersht, Enzyme Structure and Mechanism, 2nd ed., W. H. Freeman, New York, 1985. 

The overall reaction stoichiometry having been established by conventional methods, the first task of chemical kinetics is essentially the qualitative one of establishing the kinetic scheme in other words, the overall reaction is to be decomposed into its elementary reactions. This is not a trivial problem, nor is there a general solution to it. Much of Chapter 3 deals with this issue. At this point it is sufficient to note that evidence of the presence of an intermediate is often critical to an efficient solution. Modem analytical techniques have greatly assisted in the detection of reactive intermediates. A nice example is provided by a study of the pyridine-catalyzed hydrolysis of acetic anhydride. Other kinetic evidence supported the existence of an intermediate, presumably the acetylpyridinium ion, in this reaction, but it had not been detected directly. Fersht and Jencks observed (on a time scale of tenths of a second) the rise and then fall in absorbance of a solution of acetic anhydride upon treatment with pyridine. This requires that the overall reaction be composed of at least two steps, and the accepted kinetic scheme is as follows. 

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