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Endogenous substrates for protein kinases in Coprinus macrorhizus
Abstract:
The phosphorylation of cellular proteins in mycelia in strain Fisc in Coprinus macrorhizus was examined. The phosphorylation of two proteins, Protein A and B, was stimulated by cyclic AMP in the presence of Mg2+, and that of one protein, Protein C, was inhibited by cyclic AMP. The molecular weight of these proteins was determined, by gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), to be 64000 (Protein A), 46000 (Protein B), and 18000 (Protein C), respectively. These proteins were quickly phosphorylated and the phosphorylation reached to the maximal levels in 5 min. The concentration of cyclic AMP required for the half-maximal stimulation of phosphorylation of Protein A and B, and for the half-maximal inhibition of phosphorylation of Protein C was approx. 1.0 x 10(-7) M. Cyclic GMP and cyclic IMP were slightly effective for stimulation and inhibition of these proteins.
Insights
Cyclic AMP influences protein phosphorylation in Coprinus macrorhizus mycelia. It stimulates two proteins (A and B) and inhibits one (Protein C), with rapid phosphorylation observed.
Area of Science:
- Biochemistry
- Mycology
- Cellular Biology
Background:
- Cellular signaling pathways are crucial for fungal development.
- Protein phosphorylation is a key regulatory mechanism in cellular processes.
Purpose of the Study:
- To investigate the effect of cyclic AMP on protein phosphorylation in Coprinus macrorhizus mycelia.
- To identify and characterize proteins whose phosphorylation is modulated by cyclic AMP.
Main Methods:
- Mycelia of Coprinus macrorhizus (strain Fisc) were subjected to varying cyclic AMP concentrations.
- Protein phosphorylation levels were assessed using gel electrophoresis in the presence of sodium dodecyl sulfate (SDS).
- Molecular weights of affected proteins were determined.
Main Results:
- Cyclic AMP (cAMP) stimulated the phosphorylation of two proteins (Protein A, MW 64,000; Protein B, MW 46,000).
- Cyclic AMP inhibited the phosphorylation of one protein (Protein C, MW 18,000).
- Phosphorylation reached maximal levels within 5 minutes, with half-maximal effects at approximately 1.0 x 10(-7) M cAMP. Cyclic GMP and cyclic IMP showed minimal effects.
Conclusions:
- Cyclic AMP plays a significant role in regulating protein phosphorylation in Coprinus macrorhizus.
- Specific proteins (A, B, and C) are direct or indirect targets of cAMP-mediated signaling.
- The findings provide insights into the molecular mechanisms governing fungal growth and development.