Discovery of ligands for beta gamma subunits from phage-displayed peptide libraries.
We have described a method using polyvalent peptide display on filamentous phage that can be used to identify ligands that bind to G protein beta gamma subunits. Also described is how to construct phage that have known beta gamma binding sequences fused to the coat protein to allow a competition analysis to be performed. Once selected or constructed, these phage-bearing beta gamma-binding peptides are powerful tools for mapping interaction sites for beta gamma binding proteins and can be used to begin to dissect the unique modes of binding for individual beta gamma subunit-regulated effectors.
Smrcka
Alan V
AV
Department of Pharmacology and Physiology, University of Rochester, School of Medicine and Dentistry, Rochester, New York 14642, USA.
Scott
Jamie K
JK
D000595:Amino Acid Sequence; D000818:Animals; D017104:Bacteriophage M13 / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D020962:Heterotrimeric GTP-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D019151:Peptide Library; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping / Q000379:methods; D018411:Spodoptera; D014162:Transfection / Q000379:methods
D019151:Peptide Library; D021122:Protein Subunits; D011994:Recombinant Proteins; D020962:Heterotrimeric GTP-Binding Proteins