title: A luciferase-based method for assay of 5'-adenylylsulfate reductase.
abstract: A luciferase-based method was developed for measurement of 5'-adenylylsulfate (APS) reductase (APR), an enzyme of the reductive sulfate assimilation pathway in prokaryotes and plants. APR catalyzes the two-electron reduction of APS and forms sulfite and adenosine 5'-monophospahate (AMP). The luciferase-based assay measures AMP production using an enzyme-coupled system that generates luminescence. The method is shown to provide an accurate measurement of APR kinetic properties and can be used for both endpoint and continuous assays. APR activity can be measured from pure enzyme preparations as well as from crude protein extracts of tissues. In addition, the assay is ideally suited to high-throughput sample analysis of APR activity in a microtiter dish format. The method adds new capability to the study of the biochemistry and physiology of APR.
authors.affiliation: Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, USA; Institute of Maize Research, Key Laboratory of Biology and Genetic Improvement of Maize in the Southwest Region, Ministry of Agriculture, Sichuan Agricultural University, Chengdu 611130, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Xiang
authors.affiliation: Institute of Maize Research, Key Laboratory of Biology and Genetic Improvement of Maize in the Southwest Region, Ministry of Agriculture, Sichuan Agricultural University, Chengdu 611130, China.
authors.forename: Guangtang
authors.initials: G
authors.lastname: Pan
authors.affiliation: Institute of Maize Research, Key Laboratory of Biology and Genetic Improvement of Maize in the Southwest Region, Ministry of Agriculture, Sichuan Agricultural University, Chengdu 611130, China.
authors.forename: Tingzhao
authors.initials: T
authors.lastname: Rong
authors.affiliation: Department of Biological Sciences, Lehman College, City University of New York, Bronx, NY 10468, USA.
authors.forename: Zhi-Liang
authors.initials: ZL
authors.lastname: Zheng
authors.affiliation: Department of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, USA. Electronic address: leustek@aesop.rutgers.edu.
authors.forename: Thomas
authors.initials: T
authors.lastname: Leustek
mesh_terms: D000249:Adenosine Monophosphate / Q000096:biosynthesis; D000818:Animals; D057075:Enzyme Assays / Q000379:methods*; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism*; D008163:Luminescent Measurements; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000378:metabolism*; D044445:Ulva / Q000201:enzymology; D003313:Zea mays / Q000201:enzymology
chemical_list: D000249:Adenosine Monophosphate; D008156:Luciferases; D050862:Oxidoreductases Acting on Sulfur Group Donors; C019618:adenylylsulfate reductase
keywords: Adenosine 5′-monophosphate; Adenosine 5′-phosphosulfate; Luciferase; Reductase; Sulfate assimilation