title: Assay Methods for ACS Activity and ACS Phosphorylation by MAP Kinases In Vitro and In Vivo.
abstract: Ethylene, a gaseous phytohormone, has profound effects on plant growth, development, and adaptation to the environment. Ethylene-regulated processes begin with the induction of ethylene biosynthesis. There are two key steps in ethylene biosynthesis. The first is the biosynthesis of 1-aminocyclopropane-1-carboxylic acid (ACC) from S-Adenosyl-Methionine (SAM), a common precursor in many metabolic pathways, which is catalyzed by ACC synthase (ACS). The second is the oxidative cleavage of ACC to form ethylene under the action of ACC oxidase (ACO). ACC biosynthesis is the committing and generally the rate-limiting step in ethylene biosynthesis. As a result, characterizing the cellular ACS activity and understanding its regulation are important. In this chapter, we detail the methods used to measure, (1) the enzymatic activity of both recombinant and native ACS proteins, and (2) the phosphorylation of ACS protein by mitogen-activated protein kinases (MAPKs) in vivo and in vitro.
authors.affiliation: College of Life Sciences, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, 010018, P. R. China.
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Han
authors.affiliation: College of Life Sciences, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, 010018, P. R. China. liguojing@imau.edu.cn.
authors.forename: Guojing
authors.initials: G
authors.lastname: Li
authors.affiliation: Division of Biochemistry, Interdisciplinary Plant Group, and Bond Life Sciences Center, University of Missouri, 371G Bond Life Sciences Center, 117 Schweitzer Hall, Columbia, MO, 65211, USA. zhangsh@missouri.edu.
authors.forename: Shuqun
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000598:Amino Acids, Cyclic / Q000378:metabolism; D001681:Biological Assay / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005030:Ethylenes / Q000096:biosynthesis; D047468:Immunoprecipitation / Q000379:methods; D008190:Lyases / Q000235:genetics / Q000378:metabolism*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D010766:Phosphorylation; D010937:Plant Growth Regulators / Q000096:biosynthesis; D011994:Recombinant Proteins; D012588:Scintillation Counting / Q000379:methods
chemical_list: D000598:Amino Acids, Cyclic; D005030:Ethylenes; D010937:Plant Growth Regulators; D011994:Recombinant Proteins; C023863:1-aminocyclopropane-1-carboxylic acid; C036216:ethylene; D020928:Mitogen-Activated Protein Kinases; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase
keywords: 1-aminocyclopropane-1-carboxylic acid (ACC); ACC synthase (ACS); ACS activity assay; Mitogen-activated protein kinase (MAPK); Phosphorylation assay