Site-directed mutagenesis, catalytic antibodies

In the last 5 years, catalytic antibodies have been generated for several reaction types, including the various types of hydrolysis, transesterification, amide bond formation, /3-elimination, cycloreversion, transacylation, redox reactions, E-Z isomerization, epoxidation, and Diels-Alder reactions. For more information on these and other recent developments, such as semi-synthetic antibodies, site-directed mutagenesis, and the bait-and-switch strategy, the reader should consult the appropriate authorities (Schultz, 1988, 1989a,b Benkovic et al., 1990 Janda et al., 1990, 1991 Janjic and Tramontano, 1990 Lerner et al., 1991). 

Control in biocatalytic systems comes from such new techniques as directed evolution [52], the formation of catalytic antibodies using transition state templates [53], and the powerful approach of site-directed mutagenesis [54], In both heterogeneous and enzymatic catalysis, rather than take what you get, we now try to design what we want and devise clever chemical and biological approaches to get it. 

Although the site-directed mutagenesis and catalytic antibody technique are useful genetic methods, chemical modification of enzymes remains a valuable tooi for protein engineering. The latter approaches are divided into... 

If necessary, these combining sites may be equipped with an appropriately positioned catalytic amino acid side chain or a cofactor, which may be introduced either directly by site-directed mutagenesis or indirectly by chemical modification. Antibodies bind molecules ranging in size from about 6 to 34 A with association constants of M , in other words, binding is extremely strong. As with... 

Stewart, J.D., Roberts, V.A., Thomas, N.R., Getzoff, E.D., and Benkovic, S.J. (1994) Site-directed mutagenesis of a catalytic antibody An arginine and a histidine residue play key roles. Biochemistry 33, 1994, and references therein. 

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