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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Fractionation
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Cell Line
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Cyclic AMP / metabolism*
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Glyceraldehyde-3-Phosphate Dehydrogenases / chemistry*
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Glyceraldehyde-3-Phosphate Dehydrogenases / isolation & purification
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Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
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Isoenzymes / chemistry
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Isoenzymes / isolation & purification
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Isoenzymes / metabolism
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Mass Spectrometry
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Nicotiana / enzymology*
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Nucleoside-Diphosphate Kinase / chemistry*
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Nucleoside-Diphosphate Kinase / isolation & purification
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Nucleoside-Diphosphate Kinase / metabolism
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Plant Proteins / chemistry
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Plant Proteins / isolation & purification
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Plant Proteins / metabolism
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Surface Plasmon Resonance
Substances
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Isoenzymes
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Plant Proteins
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Cyclic AMP
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Glyceraldehyde-3-Phosphate Dehydrogenases
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Nucleoside-Diphosphate Kinase