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Full Description
This book describes state-of-the-art approaches for identification and functional characterization of plant peptide hormones and growth factors. Chapters guide readers through protocols for computational identification of novel signaling peptides and phytocytokines, analysis of post-translational maturation events, isolation of mature peptides from plant tissues, peptide bioassays, in vivo and in vitro analysis of peptide activity, peptide receptor identification and interaction analysis, and quantitative assessment of peptide ligand binding to cognate receptors. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Plant Peptide Hormones and Growth Factors aims to be a useful and practical guide to new researchers and experts looking to expand their technical know-how.
Contents
Bioinformatics Methods for Prediction of Gene Families Encoding Extracellular Peptides.- Identification of Bioactive Phytocytokines Using Transcriptomic Data and Plant Bioassays.- Purification of Phytaspases Using a Biotinylated Peptide Inhibitor.- Characterization of Phytaspase Proteolytic Activity Using Fluorogenic Peptide Substrates.- Characterization of prolyl-4 hydroxylase substrate specificity using Pichia pastoris as an effective eukaryotic expression system.- Extraction of Apoplastic Peptides for the Structural Elucidation of Mature Peptide Hormones in Arabidopsis.- Assaying the Effect of Peptide Treatment on H+-Pumping Activity in Plasma Membranes from Arabidopsis Seedlings.- A Seedling Growth Inhibition Assay to Measure Phytocytokine Activity.- A Trojan Horse Aproach Using Ustilago maydis to Study Apoplastic Maize (Zea mays) Peptides in situ.- Feeding Assay to Study the Effect of Phytocytokines on Direct and Indirect Defense in Maize.- A Quick Method to Analyze Peptide-Regulated Anthocyanin Biosynthesis.- Quantitative Measurement of Pattern-Triggered ROS Burst as an Early Immune Response in Tomato.- Automated Real-Time Monitoring of Extracellular pH to Assess Early Plant Defense Signaling.- Peptide-Mediated Cyclic Nucleotide Signaling in Plants: Identification and Characterization of Interactor Proteins with Nucleotide Cyclase Activity.- Detection of Ligand-Induced Receptor Kinase and Signaling Component Phosphorylation With Mn2+-Phos-Tag SDS-PAGE.- In-vivo Cross Linking of Biotinylated Peptide Ligands to Cell Surface Receptors.- Evaluation of Direct Ligand-Receptor Interactions by Photoaffinity Labeling.- Rapid Identification of Peptide-Receptor-Coreceptor Complexes in Protoplasts.- Acridinium-Based Chemiluminescent Receptor-Ligand Binding Assay for Protein/Peptide Hormones.- LuBiA (Luciferase-based Binding Assay): Glowing Peptides as Sensitive Probes to Study Ligand-Receptor Interactions.- Microscale Thermophoresis (MST) to Study Rapid Alkalinization Factor (RALF) - Receptor Interactions.- Isothermal Titration Calorimetry to Study Plant Peptide Ligand-Receptor Interactions.
