Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica.
In order to characterize a specific extracellular 21-kDa calmodulin-binding protein (named: ECBP21) from Angelica dahurica L. suspension-cultured cells, the cDNA coding for the protein has been cloned. Here, Southern blot analysis shows that there are at least two copies of ECBP21 gene in Angelica genome. Using truncated versions of ECBP21 and synthetic peptide in CaM binding assays, we mapped the calmodulin-binding domain to a 16-amino acid stretch (residues 200-215) at the C-terminal region. The ECBP21 was localized in the cell wall area by the immunogold electron microscopy and by GFP labeling method. These results define ECBP21 as a kind of an extracellular calmodulin-binding protein (CaMBP). Furthermore, using Northern blot analysis, we examined the expression dynamics of ecbp21 during the incubation of Angelica suspension-cultured cells and the treatments with some growth regulators. The above studies further provide the molecular evidence for the existence of the gene coding for extracellular CaMBPs and imply a possible role for ECBP21.
Mao
Guo-Hong
GH
Institute of Molecular Cell Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei 050016, People's Republic of China.
Hou
Li-Xia
LX
Ding
Cun-Bao
CB
Cui
Su-Juan
SJ
Sun
Da-Ye
DY
D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D029969:Angelica / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D003517:Cyclopentanes / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005875:Gibberellins / Q000494:pharmacology; D008969:Molecular Sequence Data; D054883:Oxylipins; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D020156:Salicylic Acid / Q000494:pharmacology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D003517:Cyclopentanes; D018076:DNA, Complementary; D018744:DNA, Plant; D005875:Gibberellins; D054883:Oxylipins; D010940:Plant Proteins; C011006:jasmonic acid; D000040:Abscisic Acid; D020156:Salicylic Acid