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

FEBS Letters

Witters, Erwin¹  Laukens, Kris¹  Roef, Luc¹  Van Onckelen, Harry¹  Slegers, Herman² 

[1] Laboratorium voor Plantenbiochemie en -fysiologie, Department of Biology, University of Antwerp (UIA), Universiteitsplein 1, B-2610 Antwerp, Belgium;Cellular Biochemistry, Department of Biochemistry, University of Antwerp (UIA), Universiteitsplein 1, B-2610 Antwerp, Belgium

关键词: Adenosine 3′;    5′-cyclic monophosphate;    Cell cycle;    Glyceraldehyde 3-phosphate dehydrogenase;    Nucleoside diphosphate kinase;    Tobacco bright yellow 2;    AET-cAMP;    8-(2-aminoethylthio)-cAMP;    BSA;    bovine serum albumin;    cAMP;    adenosine 3′;    5′-cyclic monophosphate;    ESI;    electrospray ionisation;    GAPDH;    glyceraldehyde 3-phosphate dehydrogenase;    MS;    mass spectrometry;    NAD+;    β-nicotinamide adenine dinucleotide;    NDPK;    nucleoside diphosphate kinase;    PKA;    protein kinase A;    TBY-2;    tobacco bright yellow 2;   

DOI  :  10.1016/S0014-5793(01)03026-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown