SUBCELLULAR DISTRIBUTION OF S-NITROSYLATED H-RAS IN DIFFERENTIATED AND UNDIFFERENTIATED PC12 CELLS DURING HYPOXIA

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BARBAKADZE TAMAR; GOLOSHVILI GALINA; NARMANIA NANA; ZHURAVLIOVA ELENE; MIKELADZE DAVID

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Journal Paper

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Journal: Cell Journal (Yakhteh) Year:2017 | Volume:19 | Issue:3 Page(s): 443-451

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Title

SUBCELLULAR DISTRIBUTION OF S-NITROSYLATED H-RAS IN DIFFERENTIATED AND UNDIFFERENTIATED PC12 CELLS DURING HYPOXIA

Author(s)

Keywords

CELL HYPOXIA

NITRIC OXIDE

H-RAS

MITOCHONDRIA

ATP

Abstract

Objective: Hypoxia or exposure to excessive reactive oxygen or nitrogen species could induce S-nitrosylation of various target proteins, including GTPases of the Ras-superfamily.Under hypoxic conditions, the Ras-protein is translocated to the cytosol and interacts with the Golgi complex, endoplasmic reticulum, MITOCHONDRIA. The mobility/translocation of Ras depend on the cells oxidative status. However, the importance of relocated Snitrosylated-H-Ras (NO-H-Ras) in proliferation/differentiation processes is not completely understood. We have determined the content of soluble- and membrane-bound-NO-HRas in differentiated (D) and undifferentiated (ND) rat pheochromocytoma (PC12) cells under hypoxic and normoxic conditions.Materials and Methods: In our experimental study, we analyzed NO-H-Ras levels under hypoxic/normoxic conditions in membrane and soluble fractions of ND and D PC12 cells with/without NITRIC OXIDE donor, sodium nitroprusside (SNP) treatment. Cells were analyzed by the S-nitrosylated kit, immunoprecipitation, and Western blot. We assessed the action of NO-H-Ras on oxidative metabolism of isolated MITOCHONDRIA by determining MITOCHONDRIAl hydrogen peroxide generation via the scopoletin oxidation method and ATPproduction as estimated by the luminometric method.Results: Hypoxia did not influence nitrosylation of soluble H-RAS in ND PC12 cells. Under hypoxic conditions, the nitrosylation of soluble-H-Ras greatly decreased in D PC12 cells. SNP didn’t change the levels of nitrosylation of soluble-H-Ras, in either hypoxic or normoxic conditions. On the other hand, hypoxia, per se, did not affect the nitrosylation of membrane-bound-H-Ras in D and ND PC12 cells. SNP-dependent nitrosylation of membrane-bound-H-Ras greatly increased in D PC12 cells. Both unmodified normal and mutated H-RAS enhanced the MITOCHONDRIAl synthesis of ATP, whereas the stimulatory effects on ATP synthesis were eliminated after S-nitrosylation of H-RAS.Conclusion: According to the results, it may be proposed that hypoxia can decrease S-nitrosylation of soluble-H-Ras in D PC12 cells and abolish the inhibitory effect of NO-HRas in MITOCHONDRIAl oxidative metabolism.

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APA: Copy

BARBAKADZE, TAMAR, GOLOSHVILI, GALINA, NARMANIA, NANA, ZHURAVLIOVA, ELENE, & MIKELADZE, DAVID. (2017). SUBCELLULAR DISTRIBUTION OF S-NITROSYLATED H-RAS IN DIFFERENTIATED AND UNDIFFERENTIATED PC12 CELLS DURING HYPOXIA. CELL JOURNAL (YAKHTEH), 19(3), 443-451. SID. https://sid.ir/paper/701444/en

Vancouver: Copy

BARBAKADZE TAMAR, GOLOSHVILI GALINA, NARMANIA NANA, ZHURAVLIOVA ELENE, MIKELADZE DAVID. SUBCELLULAR DISTRIBUTION OF S-NITROSYLATED H-RAS IN DIFFERENTIATED AND UNDIFFERENTIATED PC12 CELLS DURING HYPOXIA. CELL JOURNAL (YAKHTEH)[Internet]. 2017;19(3):443-451. Available from: https://sid.ir/paper/701444/en

IEEE: Copy

TAMAR BARBAKADZE, GALINA GOLOSHVILI, NANA NARMANIA, ELENE ZHURAVLIOVA, and DAVID MIKELADZE, “SUBCELLULAR DISTRIBUTION OF S-NITROSYLATED H-RAS IN DIFFERENTIATED AND UNDIFFERENTIATED PC12 CELLS DURING HYPOXIA,” CELL JOURNAL (YAKHTEH), vol. 19, no. 3, pp. 443–451, 2017, [Online]. Available: https://sid.ir/paper/701444/en

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