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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Acknowledgements
K. Hertveldt holds a postdoctoral position of the ``Fonds voor Wetenschappelijk Onderzoek'' (FWO)-Vlaanderen. This work was financially supported by FWO grants G017297N and G0114-01, EU contract BIO4-CT97-2294B4 and the Research Council of the K.U.Leuven (OT/98/20 en OT/04/30).
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Hertveldt, K., Robben, J. & Volckaert, G. Whole Genome Phage Display Selects for Proline-rich Boi Polypeptides against Bem1p. Biotechnol Lett 28, 1233–1239 (2006). https://doi.org/10.1007/s10529-006-9082-y
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DOI: https://doi.org/10.1007/s10529-006-9082-y