Computer-aided design of a PDZ domain to recognize new target sequences |
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Authors: | Reina Jose Lacroix Emmanuel Hobson Scott D Fernandez-Ballester Gregorio Rybin Vladimir Schwab Markus S Serrano Luis Gonzalez Cayetano |
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Institution: | Cell Biology and Biophysics Program, EMBL, 69117 Heidelberg, Germany. |
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Abstract: | PDZ domains are small globular domains that recognize the last 4-7 amino acids at the C-terminus of target proteins. The specificity of the PDZ-ligand recognition is due to side chain-side chain interactions, as well as the positioning of an alpha-helix involved in ligand binding. We have used computer-aided protein design to produce mutant versions of a Class I PDZ domain that bind to novel Class I and Class II target sequences both in vitro and in vivo, thus providing an alternative to primary antibodies in western blotting, affinity chromatography and pull-down experiments. Our results suggest that by combining different backbone templates with computer-aided protein design, PDZ domains could be engineered to specifically recognize a large number of proteins. |
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