Whole genome phage display selects for proline-rich Boi polypeptides against Bem1p.
Interaction selection by biopanning from a fragmented yeast proteome displayed on filamentous phage particles was successful in identifying proline-rich fragments of Boi1p and Boi2p. These proteins bind to the second "src homology region 3'' (SH3) domain of Bem1p, a protein of Saccharomyces cerevisiae involved in bud formation. Target Bem1p was a doubly-tagged recombinant, Bem1([Asn142-Ile551]), which strongly interacts in ELISA with a C-terminal 75 amino acids polypeptide from Cdc24p exposed on phage. The whole yeast genomic display library contained approximately 7.7 x 10(7) independent clones of sheared S. cerevisiae genomic DNA fused to a truncated M13 gene III. This study corroborates the value of fragmented-proteome display to identify strong and direct interacting protein modules.
Hertveldt
Kirsten
K
Division of Gene Technology, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, Leuven B-3001, Belgium. kirsten.hertveldt@biw.kuleuven.be
Robben
Johan
J
Volckaert
Guido
G
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry*; D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism; D016681:Genome, Fungal; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011392:Proline / Q000737:chemistry; D017434:Protein Structure, Tertiary; D040901:Proteomics / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; C099888:BOI1 protein, S cerevisiae; C073594:CDC24 protein, S cerevisiae; D018797:Cell Cycle Proteins; D020662:Guanine Nucleotide Exchange Factors; D019151:Peptide Library; D010455:Peptides; D029701:Saccharomyces cerevisiae Proteins; C073129:BEM1 protein, S cerevisiae; D011392:Proline