Genomes to Life Contractor-Grantee Workshop III
February 6-9, 2005, Washington, D.C.
Genomics:GTL Program Projects
Sandia National Laboratories
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Selection of Ligands by Panning of Phage Display Peptide Libraries Reveals Potential Partners for TPR Domain and rbcS in Synechococcus WH8102
Zhaoduo Zhang* (zzhang@sandia.gov), Arlene D. Gonzales, Todd W. Lane, and Anthony Martino
Sandia National Laboratories, Livermore, CA
One of the goals of functional genomics is the identification of reliable protein interaction partners. The oceanic cyanobacterium Synechococcus WH8102 is an abundant marine microorganism important to global CO2 fixation. We have cloned, expressed and purified two TPR domains of a conserved hypothetical protein and the RuBiscO small subunit protein rbcS from Synechococcus WH8102. After immobilizing TPR domains and rbcS, selection of ligands were carried out by panning of two phage libraries displayed random peptides. Peptides specifically binding to TPR domain or rbcS were selected and enriched after three panning processes from a 7-mer and a 12-mer library. A sequence of three amino acids TPR or TPS forms a consensus peptide specific for TPR domains, and APL or APR forms a consensus specific for rbcS. The binding of clones to the target protein was further confirmed by ELISA assay. Peptides specifically binding to rbcS were found in carboxysome protein ccmK2, orfA, csoS3, csoS2 and rbcL, potential partners for rbcS.
* Presenting author
