Mapping protein-ligand interactions using whole genome phage display libraries.
The function of many genes cannot be deduced from sequence similarity, and biochemical methods are usually required. Whole genome sequences can be thought of as not only a set of genes but also collections of functional domains. These domains can be studied by affinity methods whereby identification of the ligand can provide information on biochemical function. To take advantage of this method, one must express all functional domains in a form suitable for affinity studies. Phage display technology provides a means for accomplishing this. The pJuFo phage display system, based on the interaction between the leucine zippers Jun and Fos, has been modified and used to create a genomic phage display library from Escherichia coli MG1655. The system has been tested by using the library to map the dominant binding epitopes for an anti-RecA protein polyclonal antibody sera. This methodology provides a general biochemical approach to functional analysis of protein-ligand interactions on a genomewide basis.
Palzkill
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T
Department of Microbiology and Immunology, Baylor College of Medicine, Houston, TX 77030, USA. timothyp@bcm.tmc.edu
Huang
W
W
Weinstock
G M
GM
D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D001435:Bacteriophages / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D016762:Genes, fos / Q000235:genetics; D015698:Genomic Library; D008024:Ligands; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding; D011938:Rec A Recombinases / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013728:Terminator Regions, Genetic / Q000235:genetics
D000911:Antibodies, Monoclonal; D004269:DNA, Bacterial; D000939:Epitopes; D008024:Ligands; D011993:Recombinant Fusion Proteins; D011938:Rec A Recombinases