Dual assay for MCLV3 activity reveals structure-activity relationship of CLE peptides.
The dodecapeptide MCLV3 is a functional peptide, derived from the CLV3 precursor protein, which is a candidate ligand of the CLV1/CLV2 receptor complex that restricts the stem cell population in the shoot apical meristem (SAM). MCLV3 can induce shoot and root meristem consumption, the typical phenotype of transgenic plants overexpressing CLV3. We investigated the bioactivities of a series of alanine-substituted MCLV3 and related peptides on the root growth of Arabidopsis. The structure-activity relationship (SAR) of MCLV3 had high similarity with that of tracheary element differentiation inhibitory factor (TDIF). We also evaluated the binding activities of the peptides by a competitive receptor binding assay using tritiated MCLV3 and the membrane fraction of a tobacco BY-2 cell line overexpressing the MCLV3 ectodomain. This dual assay, combining a biological and receptor binding assay for evaluating the activities of MCLV3-related peptides, uncovered the SAR of MCLV3, and indicated that the terminal residues play critical roles in exerting its activity and are important for specific binding to the receptor, CLV1.
Kondo
Tatsuhiko
T
Graduate School of Bioagricultural Science, Nagoya University, Chikusa, Nagoya, Japan.
Nakamura
Touko
T
Yokomine
Kenjiro
K
Sakagami
Youji
Y
D000409:Alanine / Q000737:chemistry / Q000378:metabolism; D019943:Amino Acid Substitution; D017360:Arabidopsis / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D001667:Binding, Competitive; D008024:Ligands; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D018517:Plant Roots / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D011869:Radioligand Assay; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D013329:Structure-Activity Relationship
C118167:AT2G27250 protein, Arabidopsis; D029681:Arabidopsis Proteins; D008024:Ligands; C533209:MCLV3 peptide, Arabidopsis; D009842:Oligopeptides; C532424:tracheary element differentiation inhibitory factor, Arabidopsis; D020794:Receptor Protein-Tyrosine Kinases; C118166:CLV1 protein, Arabidopsis; D017346:Protein Serine-Threonine Kinases; D000409:Alanine