Increased proteolysis of oxidatively damaged hemoglobin in erythrocyte lysates in diabetes mellitus

Raghothama, Chaerkadi; Rao, Pragna

doi:10.1016/0009-8981(94)90028-0

Cited by 12 publications

(10 citation statements)

References 20 publications

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“…This indicates that diabetes, even when under moderate control, presents a severe insult of protein modification. Increased proteolysis has been found in RBCs and muscle wasting in diabetes [37,38]. If proteolysis is decreased [39], this may also lead to increased tissue protein AGE residues.…”

Section: Discussionmentioning

confidence: 99%

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Degradation products of proteins damaged by glycation, oxidation and nitration in clinical type 1 diabetes

et al. 2005

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Aims/hypothesis: Hyperglycaemia in diabetes is associated with increased glycation, oxidative stress and nitrosative stress. Proteins modified consequently contain glycation, oxidation and nitration adduct residues, and undergo cellular proteolysis with release of corresponding free adducts. These free adducts leak into blood plasma for eventual renal excretion. The aim of this study was to perform a comprehensive quantitative analysis of protein glycation, oxidation and nitration adduct residues in plasma protein and haemoglobin as well as of free adducts in plasma and urine to quantify increased protein damage and flux of proteolytic degradation products in diabetes. Methods: Type 1 diabetic patients (n=21) and normal healthy control subjects (n=12) were studied. Venous blood samples, with heparin anticoagulant, and 24-h urine samples were taken. Samples were analysed for protein glycation, oxidation and nitration adducts by a quantitative comprehensive screening method using liquid chromatography with triple quadrupole mass spectrometric detection. Results: In type 1 diabetic patients, the concentrations of protein glycation, oxidation and nitration adduct residues increased up to three-fold in plasma protein and up to one-fold in haemoglobin, except for decreases in pentosidine and 3-nitrotyrosine residues in haemoglobin when compared with normal control subjects. In contrast, the concentrations of protein glycation and oxidation free adducts increased up to ten-fold in blood plasma, and urinary excretion increased up to 15-fold in diabetic patients. Conclusions/interpretation: We conclude that there are profound increases in proteolytic products of glycated and oxidised proteins in diabetic patients, concurrent with much lower increases in protein glycation and oxidation adduct residues.

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Section: Discussionmentioning

confidence: 99%

“…This is exacerbated by low glyceraldehyde-3-phosphate dehydrogenase activity [37], particularly when inhibited by poly(ADP ribose) polymerase [44]. Ketone body metabolism may also be involved [45], which may explain why type 1 diabetic patients suffer the highest levels of methylglyoxal [46].…”

Section: Discussionmentioning

confidence: 99%

Degradation products of proteins damaged by glycation, oxidation and nitration in clinical type 1 diabetes

et al. 2005

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“…Cell-free extracts were prepared by lysing the erythrocytes in 1.5 volumes of freshly prepared 1 mM DL-dithiothreitol (DTT) (13). Intact cells and membranes were removed by centrifugation (16,000 x g for 20 minutes).…”

Section: Preparation Of Erythrocyte Cell-free Extractmentioning

confidence: 99%

“…Estimation of erythrocyte proteolytic activity (13) The erythrocyte contains several proteolytic enzymes, some of which are known to degrade oxidatively damaged hemoglobin. Under the experimental conditions used in this study, when a sample of erythrocyte lysate is incubated with phenylhydrazinetreated hemoglobin at 37°C, the enzymes in the erythrocyte degrade oxidatively damaged hemoglobin and simultaneously any other oxidant damaged protein present in the erythrocyte lysate.…”

Section: Preparation Of Erythrocyte Cell-free Extractmentioning

confidence: 99%

Maternal and fetal indicators of oxidative stress during intrauterine growth retardation (IUGR)

Kamath

Rao

Kamath

et al. 2006

Indian J Clin Biochem

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The present study demonstrates the possibility of increased lipid peroxidation and protein oxidation in both maternal and fetal erythrocytes as markers of oxygen radical activity during intrauterine growth retardation. The erythrocyte MDA levels were significantly elevated in mothers of IUGR babies when compared to controls (p<0.01). The endogenous protein damage due to oxidative stress was significantly higher in IUGR mothers when compared to controls (p<0.05). Similarly the proteolytic activity in erythrocyte lysates against oxidatively damaged hemoglobin was significantly increased in mothers of IUGR babies compared to controls (p<0.001).In fetuses born with IUGR, both lipid peroxidation and proteolytic activity were significantly increased when compared to normal newborns (p<0.01).The result of this study indicates that oxidative stress was induced both in IUGR babies and their mothers which is manifested as increased lipid peroxidation and protein oxidant damage.

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“…s free extracts were prepared by lysing the erythrocytes in 1.5 volumes of freshly prepared 1 mM DL-dithiothreitol (DTT) (12). Intact cells and membranes were removed by centdfugation (16,000 x g for 20 minutes).…”

Section: Preparation Of Oxidatively Damagedmentioning

confidence: 99%

Maternal and fetal indicators of oxidative stress in various obstetric complications

Rao

Kamath

Raghothama

et al. 2003

Indian J Clin Biochem

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The present study demonstrates the incidence of increased lipid peroxidation and protein oxidation in both maternal and fetal erythrocytes as markers of oxygen radical activity in different complications of pregnancy. In fetuses born at~er premature rupture of membranes, lipid peroxidation was significantly elevated as indicated by increased malondialdehyde levels (p<0.05) as compared to controls. Proteolytic activity in the erythrocytes of mothers in this group was also significantly high(p<0.01 ). In patients delivered by lower segment cesarian section, lipid peroxidation and proteolytic activity in maternal erythrocytes were significantly high (p<0.05 and p<0.001 respectively). In patients with prolonged second stage of labour, lipid peroxidation and proteolytic activity in maternal erythrocytes was significantly higher than in controls(p<0.001 and p<0o05 respectively). In this group, endogenous protein damage due to oxidative stress was significantly high both in the mother and the fetus (p<0.001 and p<0.05 respectively).

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Increased proteolysis of oxidatively damaged hemoglobin in erythrocyte lysates in diabetes mellitus

Cited by 12 publications

References 20 publications

Degradation products of proteins damaged by glycation, oxidation and nitration in clinical type 1 diabetes

Degradation products of proteins damaged by glycation, oxidation and nitration in clinical type 1 diabetes

Maternal and fetal indicators of oxidative stress during intrauterine growth retardation (IUGR)

Maternal and fetal indicators of oxidative stress in various obstetric complications

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