Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / chemistry*
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Amino Acid Sequence
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Cell Cycle Proteins / metabolism
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Genome, Fungal
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Guanine Nucleotide Exchange Factors / metabolism
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Mass Spectrometry
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Molecular Sequence Data
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Peptide Library*
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Peptides / chemistry*
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Proline / chemistry
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Protein Structure, Tertiary
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Proteomics / methods
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / chemistry*
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Saccharomyces cerevisiae Proteins / metabolism
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Sequence Homology, Amino Acid
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src Homology Domains
Substances
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Adaptor Proteins, Signal Transducing
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BOI1 protein, S cerevisiae
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CDC24 protein, S cerevisiae
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Cell Cycle Proteins
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Guanine Nucleotide Exchange Factors
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Peptide Library
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Peptides
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Saccharomyces cerevisiae Proteins
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BEM1 protein, S cerevisiae
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Proline