title: Identification of target ligands of CORYNE in Arabidopsis by phage display library.
abstract: CORYNE (CRN) plays important roles in stem cell division and differentiation of shoot apical meristem (SAM) in Arabidopsis thaliana. The cytoplasmic kinase domain of CRN has been cloned and expressed in Escherichia coli, and further purified by two consecutive steps of affinity chromatography. By using this purified CRN as a ligand, a 12-mer random-peptide library was used to determine the specific amino acid sequences binding with the recombinant CRN molecule. After four rounds of biopanning, positive phage clones were isolated and sequenced, and further tested by enzyme linked immunosorbent assay for their binding ability and specificity. Two positive clones that specifically bind to the intracellular protein kinase domain of CRN have been identified. Alignment of these peptides and the kinase-associated protein phosphatase (KAPP) shows high similarity, indicating that KAPP might interact with the cytoplasmic kinase domain of CRN and negatively regulate the CLV signal. Our current study would be helpful to better understand the CLV3 signal pathway.
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China. hengzhao2000@gmail.com
authors.forename: Heng
authors.initials: H
authors.lastname: Zhao
authors.forename: Shuzhen
authors.initials: S
authors.lastname: Li
authors.forename: Jiping
authors.initials: J
authors.lastname: Sheng
authors.forename: Lin
authors.initials: L
authors.lastname: Shen
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Yang
authors.forename: Bin
authors.initials: B
authors.lastname: Yao
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism*; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D005822:Genetic Vectors / Q000235:genetics; D008024:Ligands; D008969:Molecular Sequence Data; D019151:Peptide Library; D010957:Plasmids / Q000235:genetics; D017346:Protein Serine-Threonine Kinases; D017434:Protein Structure, Tertiary; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D020539:Sequence Analysis, Protein
chemical_list: D029681:Arabidopsis Proteins; D008024:Ligands; D019151:Peptide Library; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D020794:Receptor Protein-Tyrosine Kinases; C548152:CORYNE protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases