Characterisation and cloning of a calmodulin-like domain protein kinase from Chlamydomonas moewusii (Gerloff).
Calcium-stimulated protein kinase activity in the flagella of the green alga Chlamydomonas moewusii (Gerloff) was characterised. Using SDS-PAGE and an on-blot phosphorylation assay, a 65-kDa protein was identified as the major calcium-stimulated protein kinase. Its activity was directly stimulated by calcium, a characteristic of the calmodulin-like domain protein kinases (CDPKs). Monoclonal antibodies raised against the CDPK alpha from soybean cross-reacted with the 65-kDa protein in the flagella, and also with other proteins in the flagellum and cell body. The same monoclonal antibodies were used to screen a C. moewusii cDNA expression library in order to isolate CDPK cDNAs from C. moewusii. The CCK1 cDNA encodes a protein with a kinase and calmodulin-like domain linked by a junction domain typical of CDPKs. From Southern analyses, evidence was obtained for a CDPK gene family in C. moewusii and C. reinhardtii.
Siderius
M
M
Institute for Molecular Cell Biology, Biocentrum Amsterdam, University of Amsterdam, The Netherlands.
Henskens
H
H
Porto-leBlanche
A
A
van Himbergen
J
J
Musgrave
A
A
Haring
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000494:pharmacology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics*; D002696:Chlamydomonas / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D003470:Culture Media / Q000494:pharmacology; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*
D003470:Culture Media; D010940:Plant Proteins; C107257:CCK1 protein, Chlamydomonas moewusii; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium