Mechanical perturbations trigger endothelial nitric oxide synthase activity in human red blood cells
| Autor: | Jyotirmaya Behera, B. Mohammed Jaffar Ali, Uma Maheswari Balaguru, Venkatesan Saravanakumar, Rajendran Kadarkarai Raj, Vinoth Rajendran, Suvro Chatterjee, Venil N. Sumantran, Yogarajan Shathya, Shunmugan Nagarajan |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Erythrocytes Nitric Oxide Synthase Type III Gene Expression Nitric Oxide Mechanotransduction Cellular Article Cell Line Nitric oxide 03 medical and health sciences Enzyme activator chemistry.chemical_compound Western blot Cell Movement Enos Extracellular medicine Humans Amino Acid Sequence Multidisciplinary Base Sequence 030102 biochemistry & molecular biology medicine.diagnostic_test biology Nitrosylation hemic and immune systems biology.organism_classification Cell biology Enzyme Activation 030104 developmental biology Biochemistry chemistry Phosphorylation circulatory and respiratory physiology |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| DOI: | 10.1038/srep26935 |
| Popis: | Nitric oxide (NO), a vascular signaling molecule, is primarily produced by endothelial NO synthase. Recently, a functional endothelial NO synthase (eNOS) was described in red blood cells (RBC). The RBC-eNOS contributes to the intravascular NO pool and regulates physiological functions. However the regulatory mechanisms and clinical implications of RBC-eNOS are unknown. The present study investigated regulation and functions of RBC-eNOS under mechanical stimulation. This study shows that mechanical stimuli perturb RBC membrane, which triggers a signaling cascade to activate the eNOS. Extracellular NO level, estimated by the 4-Amino-5-Methylamino-2′, 7′-Difluorofluorescein Diacetate probe, was significantly increased under mechanical stimuli. Immunostaining and western blot studies confirmed that the mechanical stimuli phosphorylate the serine 1177 moiety of RBC-eNOS and activates the enzyme. The NO produced by activation of RBC-eNOS in vortexed RBCs promoted important endothelial functions such as migration and vascular sprouting. We also show that mechanical perturbation facilitates nitrosylation of RBC proteins via eNOS activation. The results of the study confirm that mechanical perturbations sensitize RBC-eNOS to produce NO, which ultimately defines physiological boundaries of RBC structure and functions. Therefore, we propose that mild physical perturbations before, after, or during storage can improve viability of RBCs in blood banks. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|