Engineering multiple properties of a protein by combinatorial mutagenesis.
A method for simultaneously engineering multiple properties of a protein, based on the observed additivity of effects of individual mutations, is presented. We show that, for the gene V protein of bacteriophage f1, effects of double mutations on both protein stability and DNA binding affinity are approximately equal to the sums of the effects of the constituent single mutations. This additivity of effects implies that it is possible to deliberately construct mutant proteins optimized for multiple properties by combination of appropriate single mutations chosen from a characterized library.
Department of Biochemistry and Molecular Biology, University of Chicago, IL 60637.
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Sandberg
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Terwilliger
D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003090:Coliphages / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D013816:Thermodynamics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D004277:DNA, Single-Stranded; D011994:Recombinant Proteins; D014764:Viral Proteins; C069558:gene V protein, Enterobacteria phage f1