Whole Genome Phage Display Selects for Proline-rich Boi Polypeptides against Bem1p |
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Authors: | Kirsten Hertveldt Johan Robben Guido Volckaert |
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Affiliation: | (1) Division of Gene Technology, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, Leuven, B-3001, Belgium;(2) Biomedical Research Institute, Limburgs Universitair Centrum and School of Life Sciences, University Hasselt, Diepenbeek, B-3590, Belgium |
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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 ~7.7 × 107 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. |
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Keywords: | Mass spectrometry Phage display Protein– protein interaction Proteomics Saccharomyces cerevisiae SH3 domain |
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