title: Cyclic AMP affinity purification and ESI-QTOF MS-MS identification of cytosolic glyceraldehyde 3-phosphate dehydrogenase and two nucleoside diphosphate kinase isoforms from tobacco BY-2 cells.
abstract: The soluble protein fraction of tobacco bright yellow 2 cells contained adenosine 3',5'-cyclic monophosphate (cAMP)-binding activity, detected with both a conventional binding assay and a surface plasmon resonance biosensor. A cAMP-agarose-based affinity purification procedure yielded three proteins which were identified by mass spectrometry as glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and two nucleoside diphosphate kinases (NDPKs). This is the first report describing an interaction between cAMP and these proteins in higher plants. Our findings are discussed in view of the reported role of the interaction of cAMP with GAPDH and NDPK in animals and yeast. In addition, we provide a rapid method to isolate both proteins from higher plants.
authors.affiliation: Laboratorium voor Plantenbiochemie en -fysiologie, Department of Biology, University of Antwerp (UIA), Universiteitsplein 1, B-2610 Antwerp, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Laukens
authors.forename: L
authors.initials: L
authors.lastname: Roef
authors.forename: E
authors.initials: E
authors.lastname: Witters
authors.forename: H
authors.initials: H
authors.lastname: Slegers
authors.forename: H
authors.initials: H
authors.lastname: Van Onckelen
mesh_terms: D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013058:Mass Spectrometry; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D020349:Surface Plasmon Resonance; D014026:Nicotiana / Q000201:enzymology*
chemical_list: D007527:Isoenzymes; D010940:Plant Proteins; D000242:Cyclic AMP; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D009701:Nucleoside-Diphosphate Kinase