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Detection of protein kinase substrates in extracts of Onchocerca volvulus
Abstract:
Extracts of Onchocerca volvulus were phosphorylated in the presence of (gamma 32P)ATP and Mg2+ by endogenous protein kinase activity and exogenous rabbit muscle catalytic sub-unit of the adenosine 3'5' monophosphate dependent protein kinase (E.C. 2.7.1.37). Sodium dodecylsulfate polyacrylamide gel electrophoretic analysis of the 32P-labelled extracts revealed at least seven (32P)-phosphoproteins with apparent Mr of 92,000; 86,000; 40,000; 27,000; 26,000; 23,000 and 17,000. The phosphorylation of the components with apparent Mr of 23,000 and 17,000 was catalysed by both endogenous and exogenous protein kinases, whereas the other components required exogenous protein kinase for their phosphorylation. The endogenous protein kinase activity was inhibited by suramin and the heat-stable protein inhibitor of the adenosine 3'5' monophosphate dependent protein kinase. The (32P)phosphoproteins identified in this investigation are probably candidate regulatory molecules in O. volvulus; though their physiological functions remain to be determined.
Insights
This study identified seven phosphoproteins in Onchocerca volvulus extracts using protein kinase activity. These proteins may play regulatory roles in the parasite, but their functions require further investigation.
Area of Science:
- Biochemistry
- Parasitology
- Molecular Biology
Background:
- Onchocerca volvulus is the causative agent of onchocerciasis, a significant human parasitic disease.
- Understanding parasite molecular mechanisms, including protein phosphorylation, is crucial for developing targeted interventions.
Purpose of the Study:
- To investigate protein kinase activity and identify phosphoproteins in Onchocerca volvulus.
- To characterize the phosphorylation patterns and potential regulatory molecules within the parasite.
Main Methods:
- Phosphorylation of O. volvulus extracts using (gamma 32P)ATP and Mg2+ with endogenous and exogenous protein kinases.
- Analysis of 32P-labelled proteins via SDS-PAGE to determine molecular weights.
- Inhibition studies using suramin and a protein kinase inhibitor to probe endogenous activity.
Main Results:
- At least seven phosphoproteins (Mr 92,000–17,000) were identified in O. volvulus extracts.
- Phosphorylation of 23,000 and 17,000 Mr proteins occurred via both endogenous and exogenous kinases.
- Other phosphoproteins required exogenous kinase activity, and endogenous kinase activity was inhibited by suramin.
Conclusions:
- Identified phosphoproteins represent potential regulatory molecules in Onchocerca volvulus.
- Further research is needed to elucidate the physiological roles of these phosphoproteins in parasite biology.