Characterization of a calcium-dependent protein kinase of tobacco leaves that is associated with the plasma membrane and is inducible by sucrose.
The plasma membrane fraction from leaves of tobacco contains a 54-kDa protein with autophosphorylation activity, and the level of this protein increases after feeding of leaves with sucrose [Ohto and Nakamura (1995) Plant Physiol. 109: 973]. The 54-kDa autophosphorylation protein could not be released from the plasma membrane by treatment with salt or alkali but could be efficiently solubilized by 1% sodium deoxycholate (NaDOC). Ion-exchange chromatography of the NaDOC-solubilized proteins in the presence of octylglucoside separated the 54-kDa autophosphorylation protein into three peaks. The autophosphorylation activity of the 54-kDa protein in peaks I and II increased after feeding of leaves with sucrose. The 54-kDa protein in the peak II fraction was enriched about 125-fold starting from the microsomal membrane fraction. The 54-kDa protein in this fraction phosphorylated histone IIIS in a calcium-dependent manner and cross-reacted with an antibody against a calcium-dependent protein kinase (CDPK) of Arabidopsis thaliana. These results suggest that the 54-kDa protein in the peak II fraction is a novel isoform of CDPK which is associated with the plasma membrane and is inducible by sucrose.
Iwata
Y
Y
Laboratory of Biochemistry, Graduate School of Bioagricultural Sciences, Nagoya University, Japan. yukikoi@nibb.ac.jp
Kuriyama
M
M
Nakakita
M
M
Kojima
H
H
Ohto
M
M
Nakamura
K
K
D017360:Arabidopsis / Q000201:enzymology; D002462:Cell Membrane / Q000201:enzymology; D002852:Chromatography, Ion Exchange; D003840:Deoxycholic Acid; D004790:Enzyme Induction; D008861:Microsomes / Q000201:enzymology; D010766:Phosphorylation; D018515:Plant Leaves; D010947:Plants, Toxic; D011494:Protein Kinases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D013395:Sucrose / Q000494:pharmacology*; D014026:Nicotiana / Q000187:drug effects / Q000201:enzymology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases
D003840:Deoxycholic Acid; D011494:Protein Kinases; D013395:Sucrose; D017871:Calcium-Calmodulin-Dependent Protein Kinases