Extracellular calmodulin-binding proteins in plants: purification of a 21-kDa calmodulin-binding protein.
A 21-kDa calmodulin (CaM)-binding protein and a 19-kDa calmodulin-binding protein were detected in 0.1 M CaCl2extracts of Angelica dahurica L. suspension-cultured cells and carrot (Daucus carota L.) suspension-cultured cells, respectively, using a biotinylated cauliflower CaM gel-overlay technique in the presence of 1 mM Ca2+. No bands, or very weak bands, were shown on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels overlayed with biotinylated cauliflower CaM when 1 mM Ca2+was replaced by 5 mM EGTA, indicating that the binding of these two CaM-binding proteins to CaM was dependent on Ca2+. Less 21-kDa CaM-binding protein was found in culture medium of Angelica dahurica suspension cells; however, a 21-kDa protein was abundant in the cell wall. We believe that the 21-kDa CaM-binding protein is mainly in the cell wall of Angelica dahurica. Based on its reaction with periodic acid-Schiff (PAS) reagent, this 21-kDa protein would appear to be a glycoprotein. The 21-kDa CaM-binding protein was purified by a procedure including Sephadex G-100 gel filtration and CM-Sepharose cation-exchange column chromatography. The purity reached 91% according to gel scanning. The purified 21-kDa CaM-binding protein inhibited the activity of CaM-dependent NAD kinase and the degree of inhibition increased with augmentation of the 21-kDa protein, which appeared to be the typical characteristic of CaM-binding protein.
Biology Department, Hebei Normal University, (050016), Shijiazhuang, Hebei Province, P.R. of China.
Tang
T
Jun
Biology Department, Hebei Normal University, (050016), Shijiazhuang, Hebei Province, P.R. of China.
Wu
W
Shupin
Biology Department, Hebei Normal University, (050016), Shijiazhuang, Hebei Province, P.R. of China.
Bai
B
Juan
Biology Department, Hebei Normal University, (050016), Shijiazhuang, Hebei Province, P.R. of China.
Sun
S
Daye
Angelica; Calmodulin-binding protein; Extracellular glycoprotein