Identification of calmodulin isoform-specific binding peptides from a phage-displayed random 22-mer peptide library.

The Journal of Biological Chemistry
Ji Young ChoiMoo Je Cho

Abstract

Plants express numerous calmodulin (CaM) isoforms that exhibit differential activation or inhibition of CaM-dependent enzymes in vitro; however, their specificities toward target enzyme/protein binding are uncertain. A random peptide library displaying a 22-mer peptide on a bacteriophage surface was constructed to screen peptides that specifically bind to plant CaM isoforms (soybean calmodulin (ScaM)-1 and SCaM-4 were used in this study) in a Ca2+-dependent manner. The deduced amino acid sequence analyses of the respective 80 phage clones that were independently isolated via affinity panning revealed that SCaM isoforms require distinct amino acid sequences for optimal binding. SCaM-1-binding peptides conform to a 1-5-10 ((FILVW)XXX(FILV) XXXX(FILVW)) motif (where X denotes any amino acid), whereas SCaM-4-binding peptide sequences conform to a 1-8-14 ((FILVW)XXXXXX(FAILVW)XXXXX(FILVW)) motif. These motifs are classified based on the positions of conserved hydrophobic residues. To examine their binding properties further, two representative peptides from each of the SCaM isoform-binding sequences were synthesized and analyzed via gel mobility shift assays, Trp fluorescent spectra analyses, and phosphodiesterase competitive inhibi...Continue Reading

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Citations

Oct 15, 2003·Trends in Plant Science·Tianbao Yang, B W Poovaiah
Jul 28, 2005·Critical Reviews in Plant Sciences·P V Sathyanarayanan, B W Poovaiah
May 3, 2005·Annual Review of Plant Biology·Nicolas BouchéHillel Fromm
Jun 18, 2005·The Plant Journal : for Cell and Molecular Biology·Maggie LevySteffen Abel
Sep 19, 2003·Biochemical and Biophysical Research Communications·Narasimhaiah SitaramRamakrishnan Nagaraj
Dec 3, 2014·Plant Science : an International Journal of Experimental Plant Biology·Hui MaYan Xiang
May 13, 2004·The Journal of Biological Chemistry·Wei HuaYing-Tang Lu
Sep 30, 2005·The Journal of Biological Chemistry·Man Soo ChoiMoo Je Cho
Aug 5, 2004·The Journal of Biological Chemistry·Tianbao YangB W Poovaiah
Jun 28, 2019·Plant Direct·Dhirodatta SenapatiSudip Chattopadhyay

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