Oxidative Inactivation of Key Metabolic Enzymes During Aging

Oliver, Cynthia N.; Fulks, Richard M.; Levine, Rodney L.; Fucci, Laura; Rivett, A. Jennifer; Roseman, J.E.; Stadtman, Earl R.

doi:10.1016/b978-0-12-601060-2.50015-6

Cited by 4 publications

(7 citation statements)

References 17 publications

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“…are associated with the accumulation of oxidized proteins [formation of protein carbonyl (RCϭO) derivatives] (1)(2)(3)(4)(5)(6)(7)(8)(9)(10). In studies to determine how protein turnover is regulated, it was demonstrated that oxidation of proteins makes them susceptible to proteolytic degradation (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21).…”

mentioning

confidence: 99%

Identification of enzymes and regulatory proteins in Escherichia coli that are oxidized under nitrogen, carbon, or phosphate starvation

Noda

Berlett

Stadtman

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Using proteomic technologies, we identified 62 proteins that are oxidized to carbonyl derivatives during growth of Escherichia coli under nitrogen starvation (NS), carbon starvation (CS), and phosphate starvation (PS) conditions. The carbonylated proteins were converted to 2,4-dinitrophenylhydrazone derivatives and these were identified using Western blotting and mass spectrometry by searching E. coli proteins in the Swiss-Prot and/or NCBI databases. Fourteen of the oxidized proteins were formed under both NS and CS conditions, and only three proteins were specifically oxidized under PS conditions. Interestingly, the carbonyl content of proteins in crude extracts of cells harvested after 48 h of stationary growth under NS and CS was significantly lower than that observed at mid-log and end-log phases of growth. In contrast, the carbonyl content of proteins in extracts of cells grown under PS conditions was fairly constant during comparable periods of growth. proteomics A ging, oxidative stress, nutritional factors, and some diseases are associated with the accumulation of oxidized proteins [formation of protein carbonyl (RCϭO) derivatives] (1-10). In studies to determine how protein turnover is regulated, it was demonstrated that oxidation of proteins makes them susceptible to proteolytic degradation (11-21). Taking advantage of proteomic technologies, we have now identified proteins that are oxidized in Escherichia coli during the exponential and stationary phases of growth under nitrogen-, carbon-, or phosphatelimiting conditions and determined the effects of starvation conditions on the ability to hydrolyze a pure preparation of oxidized glutamine synthetase. Results Protein Carbonyl (RC‫؍‬O) and Protease Activities of Cell-Free Extracts.The protein carbonyl content and abilities of crude extracts to degrade oxidized proteins of wild-type (CSH7) E. coli grown on normal medium (M9) and harvested at mid-log phase of growth and those harvested at mid-log, end-log, and after 24 h or 48 h of stationary growth in nitrogen starvation (NS), carbon starvation (CS), or phosphate starvation (PS) media are summarized in Fig. 1 Upper. It is apparent from these data that the level of RCϭO during mid-log and end-log phases of growth on NS media was appreciably higher than those observed for the corresponding phases under CS or PS conditions. However, the level of RCϭO in both NS and CS declined during 48 h of stationary growth, whereas the level in PS cells was affected little by prolonged incubation. In view of the fact that oxidized proteins are more susceptible to proteolytic degradation (13-21), we determined the effect of starvation conditions on the ability of cells to degrade oxidized 14 C-labeled glutamine synthetase. As shown in Fig. 1 Lower, the protease activity of CS cells declined significantly during the 24-48 h of stationary growth, but decreased slightly in the NS and PS cells exposed to these conditions. Identification of Proteins Oxidized During Starvation Conditions.Protein spots from 2,4-dinitrophenylhyd...

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mentioning

confidence: 99%

Identification of enzymes and regulatory proteins in Escherichia coli that are oxidized under nitrogen, carbon, or phosphate starvation

Noda

Berlett

Stadtman

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…1.1, reaction l). Moreover metal-catalyzed oxidation of proline and arginine residues of proteins leads to formation of glutamic semialdehyde derivatives of the protein, and oxidation of lysine residues leads to the formation of adipic semialdehyde derivatives (Oliver et al, 1984Amici et al, 1989;Daneshvar et al, 1997;Requena et al, 2001). Threonine residues are oxidized to 2-amino-3-keto-butyric acid derivatives (Taborsky, 1973).…”

Section: Protein Carbonylationmentioning

confidence: 97%

“…It is well established that the level of oxidized proteins increases with animal age and in the development of a number of diseases (for reviews, see Oliver et al, 1984;Takahashi and Goto, 1990;Levine and Stadtman, 1992;Stadtman, 1992Stadtman, , 1998bStadtman, , 2002Agarwal and Sohal, 1994;Butterfield and Stadtman, 1997;Berlett, 1997, 1998;Smith et al, 1999;Halliwell and Gutteridge, 1999;Levine, 2002). However, because the cellular levels of oxidized proteins are dependent on many variables, the mechanisms responsible for accumulation of oxidatively modified proteins under one condition may be very different from those in another condition.…”

Section: Accumulation Of Oxidized Proteinsmentioning

confidence: 98%

Chemical Modification of Proteins by Reactive Oxygen Species

Stadtman

Levine

2006

Redox Proteomics

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“…With regard to GS, inactivation may be mediated by a variety of enzymatic [cytochrome P-450, glucose oxidase, horseradish peroxidase, NAD(P)H oxidase, and xanthine oxidase] and nonenzymatic [ascorbic acid, dihydroxyfumaric acid, and NAD(P)H plus menadione] systems (14). Inactivation was found to result in the loss of 1 of 16 histidine residues, with the subsequent formation of 1 carbonyl group per GS subunit (8).…”

mentioning

confidence: 99%

“…Many enzymes are susceptible to oxidation by any one of several mixed-function oxidation systems. Generally, such oxidation increases the protein's vulnerability to attack by intracellular proteases (9,11,14,17). The oxidation of GS in enteric bacteria has been studied in the greatest detail.…”

mentioning

confidence: 99%

Oxygen-dependent inactivation of ribulose 1,5-bisphosphate carboxylase/oxygenase in crude extracts of Rhodospirillum rubrum and establishment of a model inactivation system with purified enzyme

Cook

Tabita

1988

J Bacteriol

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Ribulose 1,5-bisphosphate (RuBP) carboxylase/oxygenase (RuBPC/0) was inactivated in crude extracts of Rhodospirilum rubrum under atmospheric levels of oxygen; no inactivation occurred under an atmosphere of argon. RuBP carboxylase activity did not decrease in dialyzed extracts, indicating that a dialyzable factor was required for inactivation. The inactivation was inhibited by catalase. Purified RuBPC/0 is relatively oxygen stable, as no loss of activity was observed after 4 h under an oxygen atmosphere. The aerobic inactivation catalyzed by endogenous factors in crude extracts was mimicked by using a model system containing purified enzyme, ascorbate, and FeSO4 or FeCl3. Dithiothreitol was found to substitute for ascorbate in the model system. Preincubation of the purified enzyme with RuBP led to enhanced inactivation, whereas Mg2+ and HC03-significantly protected against inactivation. Unlike the inactivation catalyzed by endogenous factors from extracts of R. rubrum, inactivation in the model system was not inhibited by catalase. It is proposed that ascorbate and iron, in the presence of oxygen, generate a reactive oxygen species which reacts with a residue at the activation site, rendering the enzyme inactive.Evidence has been presented that aerobic inactivation of ribulose 1,5-bisphosphate (RuBP) carboxylase/oxygenase (RuBPC/O) in Rhodospirillum rubrum proceeds by a twostep process: the enzyme is first inactivated by an alteration or modification, and then the inactive protein is proteolytically degraded (2). This is very similar to well-described systems, such as glutamine synthetase (GS) and glutamine phosphoribosylpyrophosphate aminotransferase, in which oxidative inactivation precedes the actual degradation of the modified protein (11,21). With regard to GS, inactivation may be mediated by a variety of enzymatic [cytochrome P-450, glucose oxidase, horseradish peroxidase, NAD(P)H oxidase, and xanthine oxidase] and nonenzymatic [ascorbic acid, dihydroxyfumaric acid, and NAD(P)H plus menadione] systems (14). Inactivation was found to result in the loss of 1 of 16 histidine residues, with the subsequent formation of 1 carbonyl group per GS subunit (8). Levine has suggested that a mixed-function oxidation system generates a reactive oxygen species which reacts with a histidine residue to introduce a carbonyl moiety at the active site of the enzyme (9, 10). The oxidatively modified form of GS then becomes increasingly vulnerable to intracellular and exogenous proteases (16)(17)(18).A number of other procaryotic and eucaryotic enzymes are also susceptible to oxidative modification. Pyruvate kinase, creatine kinase, lactate dehydrogenase (14), 3-phosphoglycerate kinase (3), superoxide dismutase (5), and tyrosinase (7) are all susceptible to oxidative inactivation. Thus, loss of catalytic activity due to oxidative modification appears to be a common method of enzymatic regulation. In this study, evidence is presented that RuBPC/O is also susceptible to oxidative modification in vitro.

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Oxidative Inactivation of Key Metabolic Enzymes During Aging

Cited by 4 publications

References 17 publications

Identification of enzymes and regulatory proteins in Escherichia coli that are oxidized under nitrogen, carbon, or phosphate starvation

Identification of enzymes and regulatory proteins in Escherichia coli that are oxidized under nitrogen, carbon, or phosphate starvation

Chemical Modification of Proteins by Reactive Oxygen Species

Oxygen-dependent inactivation of ribulose 1,5-bisphosphate carboxylase/oxygenase in crude extracts of Rhodospirillum rubrum and establishment of a model inactivation system with purified enzyme

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