Proteomics screening of molecular targets of granulocyte colony stimulating factor in the mouse brain and PC12 cell line.
OBJECTIVE
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.


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.


RESULTS
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.


CONCLUSIONS
Interaction of G-CSF with these proteins can explain some of its pharmacological effects in the CNS.
Department of Pharmacology and Toxicology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Maryam
M
Ghorbani
Pharmaceutical Research Center, Department of Clinical Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Amir Hooshang
AH
Mohamadpour
Pharmaceutical Research Center, Department of Pharmacodynamy and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Soghra
S
Mehri
Pharmaceutical Research Center, Department of Medicinal Chemistry, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: abnouskh@mums.ac.ir.
Khalil
K
Abnous
Pharmaceutical Research Center, Department of Medicinal Chemistry, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: hasanzadehm@mums.ac.ir.
Mohammad
M
Hassanzadeh-Khayyat
D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D047468:Immunoprecipitation / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D058990:Molecular Targeted Therapy; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016716:PC12 Cells; D040901:Proteomics; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D018696:Neuroprotective Agents; D016179:Granulocyte Colony-Stimulating Factor; D002118:Calcium
Granulocyte colony stimulating factor; Immunoprecipitation; Molecular targets; Neuroprotection