Membrane skeletal protein S-glutathionylation and hemolysis in human red blood cells

Rossi, Ranieri; Giustarini, Daniela; Milzani, Aldo; Dalle‐Donne, Isabella

doi:10.1016/j.bcmd.2006.09.003

Cited by 30 publications

(32 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The supernatant was collected, and absorbance was measured at 540 nm with appropriate controls. The lysis percentage of RBCs was calculated from the absorbance, and a fragility curve was generated by plotting varying salt concentrations versus hemolysis (43,44).…”

Section: Methodsmentioning

confidence: 99%

“…Because UCP2 might function to reduce ROS generation, we rationalized that its deficiency may affect heme and globin biosynthesis as well the stability of mature cells (43). To determine the effect of UCP2 deficiency on heme biosynthesis, we measured the rate of iron incorporation into heme in reticulocytes isolated from UCP2 KO and WT mice 7 days after induction of anemia.…”

Section: Ucp2 Function In Erythropoiesismentioning

confidence: 99%

See 1 more Smart Citation

UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis

Elorza

Hyde

Mikkola

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

UCP2, an inner membrane mitochondrial protein, has been implicated in bioenergetics and reactive oxygen species (ROS) modulation. High levels of UCP2 mRNA were recently found in erythroid cells where UCP2 is hypothesized to function as a facilitator of heme synthesis and iron metabolism by reducing ROS production. We examined UCP2 protein expression and role in mice erythropoiesis in vivo. UCP2 was mainly expressed at early stages of erythroid maturation when cells are not fully committed in heme synthesis. Iron incorporation into heme was unaltered in reticulocytes from UCP2-deficient mice. Although heme synthesis was not influenced by UCP2 deficiency, mice lacking UCP2 had a delayed recovery from chemically induced hemolytic anemia. Analysis of progenitor cells from bone marrow and fetal liver both in vitro and in vivo revealed that UCP2 deficiency results in a significant decrease in cell proliferation at the erythropoietin-dependent phase of erythropoiesis. This was accompanied by reduction in the phosphorylated form of ERK, a ROS-dependent cytosolic regulator of cell proliferation. Analysis of ROS in UCP2 null erythroid cells revealed altered distribution of ROS, resulting in decreased cytosolic and increased mitochondrial ROS. Restoration of the cytosol oxidative state of erythroid progenitor cells by the pro-oxidant Paraquat reversed the effect of UCP2 deficiency on cell proliferation in in vitro differentiation assays. Together, these results indicate that UCP2 is a regulator of erythropoiesis and suggests that inhibition of UCP2 function may contribute to the development of anemia.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Ucp2 Function In Erythropoiesismentioning

confidence: 99%

UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis

Elorza

Hyde

Mikkola

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Several signaling proteins have been shown to undergo glutathionylation, including PTPs and transcription factors (36,63,(77)(78)(79)(80). Although the mechanism by which glutathionylation occurs is not agreed upon, this process has been shown occur in vivo in relevant cell types (81)(82)(83)(84)(85)(86). Although glutathionylation of PTP1B can be achieved in vitro with the purified enzyme through oxidation by H 2 O 2 and addition of GSH (through the sulfenic acid and sulfenyl-amide as described above) (26) (27) or by addition of a very high concentration of GSSG (63), it is not known if glutathionylation occurs enzymatically or non-enzymatically in vivo and kinetic considerations clearly argue against the latter (87).…”

Section: A Signaling Proteins In Which Critical Cysteines Are Modifiedmentioning

confidence: 99%

The chemistry of cell signaling by reactive oxygen and nitrogen species and 4-hydroxynonenal

Forman

Fukuto

Miller

et al. 2008

Archives of Biochemistry and Biophysics

219

165

View full text Add to dashboard Cite

During the past several years, major advances have been made in understanding how reactive oxygen species (ROS) and nitrogen species (RNS) participate in signal transduction. Identification of the specific targets and the chemical reactions involved still remains to be resolved with many of the signaling pathways in which the involvement of reactive species has been determined. Our understanding is that ROS and RNS have second messenger roles. While cysteine residues in the thiolate (ionized) form found in several classes of signaling proteins can be specific targets for reaction with H 2 O 2 and RNS, better understanding of the chemistry, particularly kinetics, suggests that for many signaling events in which ROS and RNS participate, enzymatic catalysis is more likely to be involved than non-enzymatic reaction. Due to increased interest in how oxidation products, particularly lipid peroxidation products, also are involved with signaling, a review of signaling by 4-hydroxy-2-nonenal (HNE) is included. This article focuses on the chemistry of signaling by ROS, RNS, and HNE and will describe reactions with selected target proteins as representatives of the mechanisms rather attempt to comprehensively review the many signaling pathways in which the reactive species are involved. Keywords signaling; glutathione; thioredoxin; oxidants; reactive oxygen species; thiols; peroxide; nitric oxide; peroxynitrite; 4-hydroxynonenal; cysteine; hydrogen peroxide; protein tyrosine phosphatase; reactive nitrogen species; eNOS; iNOS; nNOS; soluble guanylate cyclase; cGMP; tyrosine nitration; fatty acid nitration; NO-heme; NO-metal complexes; nitrite; protein kinase C; ERK; JNK; p38MAPK; tyrosine kinase receptors; calcium OVERVIEW OF SIGNALING BY REACTIVE SPECIESUnderstanding of the roles of reactive oxygen species (ROS), reactive nitrogen species (RNS) and the lipid peroxidation product, 4-hydroxy-2-nonenal (HNE) in signaling has evolved rapidly during the last decade. This has been markedly helped by identification of the specific targets in signaling pathways. In previous reviews, we defined how ROS, H 2 O 2 in particular,

show abstract

“…Additionally, the formation of glutathionyl proteins in human RBCs has been reported in in vitro studies. These studies indicated that hemoglobin, GAPDH and actin, 14) as well as spectrin and membrane protein band 4.2, 15) are glutathionylated in response to diamide or ROS. Alternatively, GSH is likely to contribute to the protection of all thiol residues in proteins against ROS in the cell.…”

Section: 2)mentioning

confidence: 99%

Disruption of Glutathione Homeostasis Causes Accumulation of <i>S</i>-Glutathionyl Proteins in Response to Exposure to Reactive Oxygen Species in Human Erythrocytes

Ogasawara

Komiyama

Funakoshi

et al. 2010

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

Proteins are susceptible to attack by reactive oxygen species (ROS). Several types of ROS-induced protein damage have been identified, including amino acid modification, cross-linking, and degradation. 1,2) Glutathione (GSH) is the major intracellular non-protein thiol compound and plays a key role in the protection of cells and tissue structures from oxidative injury. GSH can exist in a reduced form (GSH), oxidized form (GSSG), or as proteinbound GSH (GSSP). GSH acts as an intracellular antioxidant that is involved in preventing protein sulfhydryl groups from oxidizing and cross-linking. In addition, the GSH redox system functions indirectly with glutaredoxin (Grx), glucose 6-phosphate dehydrogenase (G6PDH), and glutathione reductase (GR), which maintain GSH in its reduced form. An increase in oxidative stress leads to the oxidation of GSH to GSSG and, under certain conditions, the formation of glutathionyl proteins. Various glutathionyl proteins are involved in numerous physical processes (e.g., growth, differentiation, cell cycle progression, cytoskeletal functions, and metabolism), suggesting that S-glutathionylation is a general mechanism of redox regulation. [3][4][5] Red blood cells (RBCs) have been used as a model system in numerous studies to determine the significance of the GSH-dependent redox system 6,7) because they have an efficient antioxidant defense system that prevents oxidative damage.8-10) Moreover, RBCs have high levels of GSH and Grx, which are thought to utilize the reducing power of GSH to catalyze disulfide reductions in the presence of reduced nicotinamide adenine dinucleotide phosphate (NADPH) and GR. 11)Many lines of evidence have emerged to support a fundamental role for the glutathionylation of certain proteins in normal cellular biochemical and physiological processes. It has been reported that some proteins are capable of reacting to form the glutathionyl protein. In particular, it is well known that actin 12) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) 13) are oxidized to form glutathionyl proteins. Additionally, the formation of glutathionyl proteins in human RBCs has been reported in in vitro studies. These studies indicated that hemoglobin, GAPDH and actin, 14) as well as spectrin and membrane protein band 4.2, 15) are glutathionylated in response to diamide or ROS. Alternatively, GSH is likely to contribute to the protection of all thiol residues in proteins against ROS in the cell. However, the physiological susceptibility of proteins to ROS and the direct effects of GSH on oxidative stress in human RBCs remain poorly understood. Moreover, there is little information on the significance of redox regulation that is dependent on the glutathionylation of individual proteins.The objective of this study was to identify physiologically relevant glutathionyl proteins formed in RBCs by ROS exposure. Therefore, we designed experiments to disrupt GSH homeostasis.16) We used two independent treatments aimed at decreasing the level of the reduced form of GSH concretely: 1) pre-...

show abstract

Membrane skeletal protein S-glutathionylation and hemolysis in human red blood cells

Cited by 30 publications

References 53 publications

UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis

UCP2 Modulates Cell Proliferation through the MAPK/ERK Pathway during Erythropoiesis and Has No Effect on Heme Biosynthesis

The chemistry of cell signaling by reactive oxygen and nitrogen species and 4-hydroxynonenal

Disruption of Glutathione Homeostasis Causes Accumulation of <i>S</i>-Glutathionyl Proteins in Response to Exposure to Reactive Oxygen Species in Human Erythrocytes

Contact Info

Product

Resources

About