Details
| Original language | English |
|---|---|
| Article number | 183096 |
| Journal | Biochimica et biophysica acta: Biomembranes |
| Volume | 1862 |
| Issue number | 2 |
| Early online date | 28 Oct 2019 |
| Publication status | Published - 1 Feb 2020 |
| Externally published | Yes |
Abstract
Hydrogen peroxide (H2O2) plays a central role in redox signalling and in oxidative stress-mediated cell death. It is generated through multiple mechanisms at various intracellular sites. Due to its chemical stability it can reach distant sites of action. However, its hydrophilicity can hamper lipid membrane passage. We therefore studied the kinetics of H2O2 diffusion through subcellular membranes employing the H2O2 biosensor HyPer in insulin-producing RINm5F cells. Plasma- and ER-membrane-bound HyPer sensors facing the cytosolic compartment reacted twice as fast to H2O2 compared to sensors expressed in peroxisomes and mitochondria. Overexpression of the H2O2-inactivating enzyme catalase in the ER-lumen and in the peroxisomes retarded the reaction time of HyPer, both localised within the peroxisomes as well as at the cytosolic surface of the ER. The unsaturated fatty acid oleic acid did not affect the reaction of the peroxisomal HyPer sensor to H2O2, while the saturated fatty acid palmitic acid accelerated its reaction time to H2O2 in this organelle. The results show that the plasma-, peroxisomal, and mitochondrial membrane of insulin-producing RINm5F cells are permeable for H2O2. Nonetheless, the organelle membranes retard H2O2 diffusion due to a barrier function of the lipid membrane, as documented by retarded reaction times of the intraorganellar sensors. Accelerated decomposition of H2O2 by catalase, expressed in the peroxisomes or the ER, further retarded the HyPer sensor reaction time. The results show that redox signalling and oxidative stress-mediated toxicity are crucially dependent on physicochemical membrane properties and antioxidative defence mechanisms in health and disease.
Keywords
- Biosensing Techniques, Cell Membrane/metabolism, Diffusion, Humans, Hydrogen Peroxide/metabolism, Insulin-Secreting Cells/metabolism, Kinetics, Oxidation-Reduction, Oxidative Stress, Permeability, Oxidative stress, Hydrogen peroxide, Membrane permeability, Free fatty acids, Beta-cells, Genetically encoded sensors
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biophysics
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: Biochimica et biophysica acta: Biomembranes, Vol. 1862, No. 2, 183096, 01.02.2020.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Hydrogen peroxide permeability of cellular membranes in insulin-producing cells
AU - Laporte, Anna
AU - Lortz, Stephan
AU - Schaal, Christina
AU - Lenzen, Sigurd
AU - Elsner, Matthias
N1 - Funding Information: We are grateful to Maren Böger, Britta Leß and Martin Wirth for their skilful technical assistance. This work was supported by Deutsche Forschungsgemeinschaft , GRK 1947/1 .
PY - 2020/2/1
Y1 - 2020/2/1
N2 - Hydrogen peroxide (H2O2) plays a central role in redox signalling and in oxidative stress-mediated cell death. It is generated through multiple mechanisms at various intracellular sites. Due to its chemical stability it can reach distant sites of action. However, its hydrophilicity can hamper lipid membrane passage. We therefore studied the kinetics of H2O2 diffusion through subcellular membranes employing the H2O2 biosensor HyPer in insulin-producing RINm5F cells. Plasma- and ER-membrane-bound HyPer sensors facing the cytosolic compartment reacted twice as fast to H2O2 compared to sensors expressed in peroxisomes and mitochondria. Overexpression of the H2O2-inactivating enzyme catalase in the ER-lumen and in the peroxisomes retarded the reaction time of HyPer, both localised within the peroxisomes as well as at the cytosolic surface of the ER. The unsaturated fatty acid oleic acid did not affect the reaction of the peroxisomal HyPer sensor to H2O2, while the saturated fatty acid palmitic acid accelerated its reaction time to H2O2 in this organelle. The results show that the plasma-, peroxisomal, and mitochondrial membrane of insulin-producing RINm5F cells are permeable for H2O2. Nonetheless, the organelle membranes retard H2O2 diffusion due to a barrier function of the lipid membrane, as documented by retarded reaction times of the intraorganellar sensors. Accelerated decomposition of H2O2 by catalase, expressed in the peroxisomes or the ER, further retarded the HyPer sensor reaction time. The results show that redox signalling and oxidative stress-mediated toxicity are crucially dependent on physicochemical membrane properties and antioxidative defence mechanisms in health and disease.
AB - Hydrogen peroxide (H2O2) plays a central role in redox signalling and in oxidative stress-mediated cell death. It is generated through multiple mechanisms at various intracellular sites. Due to its chemical stability it can reach distant sites of action. However, its hydrophilicity can hamper lipid membrane passage. We therefore studied the kinetics of H2O2 diffusion through subcellular membranes employing the H2O2 biosensor HyPer in insulin-producing RINm5F cells. Plasma- and ER-membrane-bound HyPer sensors facing the cytosolic compartment reacted twice as fast to H2O2 compared to sensors expressed in peroxisomes and mitochondria. Overexpression of the H2O2-inactivating enzyme catalase in the ER-lumen and in the peroxisomes retarded the reaction time of HyPer, both localised within the peroxisomes as well as at the cytosolic surface of the ER. The unsaturated fatty acid oleic acid did not affect the reaction of the peroxisomal HyPer sensor to H2O2, while the saturated fatty acid palmitic acid accelerated its reaction time to H2O2 in this organelle. The results show that the plasma-, peroxisomal, and mitochondrial membrane of insulin-producing RINm5F cells are permeable for H2O2. Nonetheless, the organelle membranes retard H2O2 diffusion due to a barrier function of the lipid membrane, as documented by retarded reaction times of the intraorganellar sensors. Accelerated decomposition of H2O2 by catalase, expressed in the peroxisomes or the ER, further retarded the HyPer sensor reaction time. The results show that redox signalling and oxidative stress-mediated toxicity are crucially dependent on physicochemical membrane properties and antioxidative defence mechanisms in health and disease.
KW - Biosensing Techniques
KW - Cell Membrane/metabolism
KW - Diffusion
KW - Humans
KW - Hydrogen Peroxide/metabolism
KW - Insulin-Secreting Cells/metabolism
KW - Kinetics
KW - Oxidation-Reduction
KW - Oxidative Stress
KW - Permeability
KW - Oxidative stress
KW - Hydrogen peroxide
KW - Membrane permeability
KW - Free fatty acids
KW - Beta-cells
KW - Genetically encoded sensors
UR - http://www.scopus.com/inward/record.url?scp=85075334608&partnerID=8YFLogxK
U2 - 10.1016/j.bbamem.2019.183096
DO - 10.1016/j.bbamem.2019.183096
M3 - Article
C2 - 31672544
VL - 1862
JO - Biochimica et biophysica acta: Biomembranes
JF - Biochimica et biophysica acta: Biomembranes
SN - 0005-2736
IS - 2
M1 - 183096
ER -