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Calcium-dependent but calmodulin-independent protein kinase from soybean. [Glycine max]

Journal Article · · Plant Physiol.; (United States)
OSTI ID:5900390
; ;
A calcium-dependent protein kinase activity from suspension-cultured soybean cells (Glycine max L. Wayne) was shown to be dependent on calcium but not calmodulin. The concentrations of free calcium required for half-maximal histone H1 phosphorylation and autophosphorylation were similar (greater than or equal to 2 micromolar). The protein kinase activity was stimulated 100-fold by greater than or equal to 10 micromolar-free calcium. When exogenous soybean or bovine brain calmodulin was added in high concentration (1 micromolar) to the purified kinase, calcium-dependent and -independent activities were weakly stimulated (less than or equal to 2-fold). Bovine serum albumin had a similar effect on both activities. The kinase was separated from a small amount of contaminating calmodulin by sodium dodecyl sulfate polyacrylamide gel electrophoresis. After renaturation the protein kinase autophosphorylated and phosphorylated histone H1 in a calcium-dependent manner. Following electroblotting onto nitrocellulose, the kinase bound /sup 45/Ca/sup 2 +/ in the presence of KCl and MgCl/sub 2/, which indicated that the kinase itself is a high-affinity calcium-binding protein. Also, the mobility of one of two kinase bands in SDS gels was dependent on the presence of calcium. Autophosphorylation of the calmodulin-free kinase was inhibited by the calmodulin-binding compound N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W-7), showing that the inhibition of activity by W-7 is independent of calmodulin. These results show that soybean calcium-dependent protein kinase represents a new class of protein kinase which requires calcium but not calmodulin for activity.
Research Organization:
Univ. of Georgia, Athens
OSTI ID:
5900390
Journal Information:
Plant Physiol.; (United States), Journal Name: Plant Physiol.; (United States) Vol. 83:4; ISSN PLPHA
Country of Publication:
United States
Language:
English

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Related Subjects

551001* -- Physiological Systems-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH ISOTOPES
ALKALINE EARTH METALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL FUNCTIONS
CALCIUM
CALCIUM 45
CALCIUM ISOTOPES
CATIONS
CHARGED PARTICLES
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
ELEMENTS
ENZYME ACTIVITY
ENZYMES
EVEN-ODD NUCLEI
FOOD
FUNCTIONS
INTERMEDIATE MASS NUCLEI
IONS
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
METALS
NUCLEI
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PHYSIOLOGY
PLANTS
PURIFICATION
RADIOISOTOPES
SOYBEANS
TRACER TECHNIQUES
TRANSFERASES
VEGETABLES