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Effects of age and calorie restriction on tryptophan nitration, protein content, and activity of succinyl-CoA:3-ketoacid CoA transferase in rat kidney mitochondria

Brégère, Catherine and Rebrin, Igor and Gallaher, Timothy K. and Sohal, Rajindar S. (2010) Effects of age and calorie restriction on tryptophan nitration, protein content, and activity of succinyl-CoA:3-ketoacid CoA transferase in rat kidney mitochondria. Free Radical Biology and Medicine, 48 (4). pp. 609-618. ISSN 0891-5849. https://resolver.caltech.edu/CaltechAUTHORS:20100304-091656483

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Abstract

This study examined the protein targets of nitration and the consequent impact on protein function in rat kidney mitochondria at 4, 13, 19, and 24 months of age. Succinyl-CoA transferase (SCOT), a rate-limiting enzyme in the degradation of ketone bodies, was the most intensely reactive protein against anti-3-nitrotyrosine antibody in rat kidney mitochondria. However, subsequent mass spectrometric and amino acid analyses of purified SCOT indicated that tryptophan 372, rather than a tyrosine residue, was the actual site of simultaneous additions of nitro and hydroxy groups. This finding suggests that identification of nitrated tyrosine residues based solely on reactivity with anti-3-nitrotyrosine antibody can be potentially misleading. Between 4 and 24 months of age, the amounts of SCOT protein and catalytic activity, expressed per milligram of mitochondrial proteins, decreased by 55 and 45%, respectively. SCOT, and particularly its nitrated carboxy-terminal region, was relatively more susceptible to in vitro proteolysis than other randomly selected kidney mitochondrial proteins. The age-related decreases in SCOT protein amount and catalytic activity were prevented by a relatively long-term 40% reduction in the amount of food intake. Loss of SCOT protein in the aged rats may attenuate the capacity of kidney mitochondria to utilize ketone bodies for energy production.


Item Type:Article
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URLURL TypeDescription
http://dx.doi.org/10.1016/j.freeradbiomed.2009.12.009 DOIArticle
Additional Information:© 2009 Elsevier Inc. Received 20 May 2009; revised 7 December 2009; accepted 10 December 2009. Available online 16 December 2009. This research was supported by Grant RO1 AG 13563 from the National Institute on Aging, National Institutes of Health. Mass spectrometric analysis was performed at the Proteomics Core Facility of the University of Southern California, School of Pharmacy. We thank Dr. Sergey Kamzalov for technical assistance in mitochondrial preparation and catalytic activity measurements.
Funders:
Funding AgencyGrant Number
NIHRO1 AG13563
Subject Keywords:Protein nitration; Tryptophan nitration; Succinyl-CoA transferase; Aging; Caloric restriction; Mitochondrial enzymes; Free radicals
Issue or Number:4
Record Number:CaltechAUTHORS:20100304-091656483
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20100304-091656483
Official Citation:Catherine Bregere, Igor Rebrin, Timothy K. Gallaher, Rajindar S. Sohal, Effects of age and calorie restriction on tryptophan nitration, protein content, and activity of succinyl-CoA:3-ketoacid CoA transferase in rat kidney mitochondria, Free Radical Biology and Medicine, Volume 48, Issue 4, 15 February 2010, Pages 609-618, ISSN 0891-5849, DOI: 10.1016/j.freeradbiomed.2009.12.009.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:17656
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:12 Mar 2010 19:03
Last Modified:03 Oct 2019 01:31

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