Phosphoproteome Profiling of SH-SY5y Neuroblastoma Cells Treated with Anesthetics: Sevoflurane and Isoflurane Affect the Phosphorylation of Proteins Involved in Cytoskeletal Regulation.
Inhalation anesthetics are used to decrease the spinal cord transmission of painful stimuli. However, the molecular or biochemical processes within cells that regulate anesthetic-induced responses at the cellular level are largely unknown. Here, we report the phosphoproteome profile of SH-SY5y human neuroblastoma cells treated with sevoflurane, a clinically used anesthetic. Phosphoproteins were isolated from cell lysates and analyzed using two-dimensional gel electrophoresis. The phosphorylation of putative anesthetic-responsive marker proteins was validated using western blot analysis in cells treated with both sevoflurane and isoflurane. A total of 25 phosphoproteins were identified as differentially phosphorylated proteins. These included key regulators that signal cytoskeletal remodeling steps in pathways related to vesicle trafficking, axonal growth, and cell migration. These proteins included the Rho GTPase, Ras-GAP SH3 binding protein, Rho GTPase activating protein, actin-related protein, and actin. Sevoflurane and isoflurane also resulted in the dissolution of F-actin fibers in SH-SY5y cells. Our results show that anesthetics affect the phosphorylation of proteins involved in cytoskeletal remodeling pathways.
Department of Food and Nutrition, Chosun University, Gwangju 61452, Korea.
Joomin
J
Lee
Department of Applied Chemistry, Dongduk Women's University, Seoul 02748, Korea.
Eunsook
E
Ahn
Department of Anesthesia and Pain, College of Medicine, Department of Anesthesia and Pain, Yonsei University, Seoul 03722, Korea.
Wyun Kon
WK
Park
Department of Applied Chemistry, Dongduk Women's University, Seoul 02748, Korea.
Seyeon
S
Park
D018685:Anesthetics, Inhalation / Q000494:pharmacology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000187:drug effects; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D047468:Immunoprecipitation; D007530:Isoflurane / Q000494:pharmacology / Q000627:therapeutic use*; D008738:Methyl Ethers / Q000494:pharmacology / Q000627:therapeutic use*; D009447:Neuroblastoma / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D000077149:Sevoflurane; D053719:Tandem Mass Spectrometry
D018685:Anesthetics, Inhalation; D008738:Methyl Ethers; D010750:Phosphoproteins; D000077149:Sevoflurane; D007530:Isoflurane