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Proteomics screening of molecular targets of granulocyte colony stimulating factor in the mouse brain and PC12 cell
Maryam Ghorbani1, Amir Hooshang Mohamadpour2, Soghra Mehri3
1Department of Pharmacology and Toxicology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Aims:
Granulocyte colony stimulating factor (G-CSF), a new neuroprotective agent, binds to its specific receptors in the brain. In this study we hypothesized that at least a part of G-CSF's neuroprotective effect may be mediated through its interaction with other proteins in the brain.
Main Methods:
Using an immunoprecipitation (IP) kit, at first the antibody of G-CSF was covalently crosslinked to protein A/G agarose. Then the mouse brain or PC12 cell lysate mixed with G-CSF was added to the agarose beads plus antibody. After immunoaffinity isolation of target proteins, gel electrophoresis was performed and protein bands were identified using MALDI-TOF/TOF and MASCOT software.
Key Findings:
Our data show that G-CSF physically binds to cellular proteins like sodium/potassium-transporting ATPase, beta actin, aldehyde dehydrogenase, regucalcin and glutathione-s-transferase. These proteins are involved in membrane transportation, cell structure, signal transduction, enzymes involve in calcium related cell signaling and redox homeostasis.
Significance:
Interaction of G-CSF with these proteins can explain some of its pharmacological effects in the CNS.
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