Protein phosphorylation/dephosphorylation in the inner membrane of potato tuber mitochondria

Struglics, A.; Fredlund, Kenneth M.; Konstantinov, Yu. M.; Allen, John F.; Møller, Ian Max

doi:10.1016/s0014-5793(00)01680-x

Cited by 43 publications

(33 citation statements)

References 20 publications

(29 reference statements)

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“…In addition, although considerable evidence has accumulated to show that oxidative stress is involved in mitochondrial pathologies, no studies have linked the effect of oxidants on tyrosine phosphorylation in OXPHOS. The aim of the first set of experiments described here was to detect tyrosine-phosphorylated protein bands in brain mitochondria using an in vitro approach with added ATP, as generally performed [3,8,10,13,16]. The results demonstrated that tyrosine kinases are active in mitochondria and phosphorylate several proteins, named A-M ( fig.…”

Section: Discussionmentioning

confidence: 92%

“…However, the regulation of oxidative phosphorylation (OXPHOS) is still unclear, and the role played by ROS and protein phosphorylation remains elusive. For example, reversible protein phosphorylation has been investigated in the mitochondria [3][4][5][6][7], and results showed the existence of a cAMP-dependent protein kinase (PKA) in the inner mitochondrial membrane and matrix fraction. Of particular interest are data showing PKA-dependent phosphorylation of the 18-kDa (AQDQ) subunit of complex I [6] and of subunit I of cytochrome c oxidase * Corresponding author.…”

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confidence: 99%

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Identification of tyrosine-phosphorylated proteins of the mitochondrial oxidative phosphorylation machinery

Augereau

Claverol

Boudes

et al. 2005

CMLS, Cell. Mol. Life Sci.

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The role of some serine/threonine kinases in the regulation of mitochondrial physiology is now well established, but little is known about mitochondrial tyrosine kinases. We showed that tyrosine phosphorylation of rat brain mitochondrial proteins was increased by in vitro addition of ATP and H2O2, and also during in situ ATP production at state 3, and maximal reactive oxygen species production. The Src kinase inhibitor PP2 decreased tyrosine phosphorylation and respiratory rates at state 3. We found that the 39-kDa subunit of complex I was tyrosine phosphorylated, and we identified putative tyrosine-phosphorylated subunits for the other complexes. We also have strong evidence that the FoF1-ATP synthase alpha chain is probably tyrosine-phosphorylated, but demonstrated that the beta chain is not. The tyrosine phosphatase PTP 1B was found in brain but not in muscle, heart or liver mitochondria. Our results suggest that tyrosine kinases and phosphatases are involved in the regulation of oxidative phosphorylation.

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Section: Discussionmentioning

confidence: 92%

mentioning

confidence: 99%

Identification of tyrosine-phosphorylated proteins of the mitochondrial oxidative phosphorylation machinery

Augereau

Claverol

Boudes

et al. 2005

CMLS, Cell. Mol. Life Sci.

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“…Incubating intact mitochondria (3)(4)(5) or submitochondrial fractions like the outer membrane (6) or the inner membrane (7)(8)(9) with [␥- 32 P]ATP gives labeling of up to 30 proteins (as detected by one-dimensional gel electrophoresis) indicating that kinases are present in these fractions and in contact with target proteins. The matrix appears to contain one of more protein phosphatases responsible for dephosphorylation of inner membrane phosphoproteins (8,9).…”

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confidence: 99%

Phosphorylation of Formate Dehydrogenase in Potato Tuber Mitochondria

Bykova¹,

Stensballe

Egsgaard³

et al. 2003

Journal of Biological Chemistry

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Two highly phosphorylated proteins were detected after two-dimensional (blue native/SDS-PAGE) gel electrophoretic separation of the matrix fraction isolated from potato tuber mitochondria. These two phosphoproteins were identified by mass spectrometry as formate dehydrogenase (FDH) and the E1␣-subunit of pyruvate dehydrogenase (PDH). Isoelectric focusing/SDS-PAGE two-dimensional gels separated FDH and PDH and resolved several different phosphorylated forms of FDH. By using combinations of matrix-assisted laser desorption/ionization mass spectrometry and electrospray ionization tandem mass spectrometry, several phosphorylation sites were identified for the first time in Reversible protein phosphorylation is one of the most important regulatory mechanisms in living cells (1), yet little is know about protein phosphorylation in mitochondria. The only well studied case is that of phosphorylation of the E1␣-subunit of the pyruvate dehydrogenase (PDH) 1 complex (2). The PDH complex contains both a kinase and a protein phosphatase, and phosphorylation inactivates the complex, which is reactivated by dephosphorylation. This means that the entry of carbon compounds from glycolysis into the citric acid cycle is turned off under conditions of high ATP concentration. Although the PDH kinase and phosphatase system is the only fully characterized system for reversible protein phosphorylation in mitochondria, there are clear indications that others are present. Incubating intact mitochondria (3-5) or submitochondrial fractions like the outer membrane (6) or the inner membrane (7-9) with [␥-32 P]ATP gives labeling of up to 30 proteins (as detected by one-dimensional gel electrophoresis) indicating that kinases are present in these fractions and in contact with target proteins. The matrix appears to contain one of more protein phosphatases responsible for dephosphorylation of inner membrane phosphoproteins (8, 9).Only eight mitochondrial phosphoproteins have been identified to date. In addition to the E1␣-subunit of PDH, these are band ␦Ј-subunits of the ATP synthase (10), HSP70 (11, 12) and MTSHP (13,14), nucleoside diphosphate kinase (15, 16), E1 of the branched-chain ␣-ketoacid dehydrogenase (2), and the 18-kDa iron protein subunit of complex I in mammalian mitochondria (17). In a very recent study (18) a further 14 mitochondrial phosphoproteins were identified, all household proteins involved in the citric acid cycle and associated reactions, the respiratory chain complexes, in heat shock proteins, and the detoxification of reactive oxygen species.Formate dehydrogenase (FDH) catalyzes the oxidation of formate to CO 2 reducing NAD ϩ to NADH in the process. Formate may have a role in biosynthesis of numerous compounds, such as in energetic metabolism and in signal transduction pathways related to stress response. The enzyme is induced in potato leaves by a variety of stresses including hypoxia (19,20). It is one of the most abundant proteins in the matrix of potato tuber mitochondria (19), but its function is unknown partly be...

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“…This high turnover confirmed the existence of an active kinase and phosphatase in the matrix. Subsequently, this approach has been used extensively in intact potato mitochondria (201), isolated potato mitochondria membranes (164,204), porcine heart mitochondria (5,6,83,161), as well as in blue native gel resolved mitochondrial complexes (161). These studies demonstrate extensive 32 P incorporation into matrix-exclusive proteins once proper controls are run for weak and tight metabolite associations (5).…”

Section: Mitochondrial Matrix Protein Kinasesmentioning

confidence: 99%

“…Thus, even though extensive phosphatase activity is present in the matrix based on 32 P turnover and quantitative MS studies (24), the phosphatases of the matrix remain obscure. Added evidence that these phosphatases exist in the aqueous phase in the matrix was the demonstration of phosphatase activity on the inner membrane by a matrix extract in potato mitochondria (204). Clearly, the protein phosphatase system within the mitochondria matrix is as ill defined as are the matrix kinase components, and a clear mechanism to explain the larger turnover of protein phosphorylation in the matrix is yet to be resolved.…”

Section: Matrix Protein Phosphatasesmentioning

confidence: 99%

Cardiac mitochondrial matrix and respiratory complex protein phosphorylation

Covián

Balaban

2012

American Journal of Physiology-Heart and Circulatory Physiology

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Covian R, Balaban RS. Cardiac mitochondrial matrix and respiratory complex protein phosphorylation. Am J Physiol Heart Circ Physiol 303: H940 -H966, 2012. First published August 10, 2012; doi:10.1152/ajpheart.00077.2012.-It has become appreciated over the last several years that protein phosphorylation within the cardiac mitochondrial matrix and respiratory complexes is extensive. Given the importance of oxidative phosphorylation and the balance of energy metabolism in the heart, the potential regulatory effect of these classical signaling events on mitochondrial function is of interest. However, the functional impact of protein phosphorylation and the kinase/phosphatase system responsible for it are relatively unknown. Exceptions include the well-characterized pyruvate dehydrogenase and branched chain ␣-ketoacid dehydrogenase regulatory system. The first task of this review is to update the current status of protein phosphorylation detection primarily in the matrix and evaluate evidence linking these events with enzymatic function or protein processing. To manage the scope of this effort, we have focused on the pathways involved in energy metabolism. The high sensitivity of modern methods of detecting protein phosphorylation and the low specificity of many kinases suggests that detection of protein phosphorylation sites without information on the mole fraction of phosphorylation is difficult to interpret, especially in metabolic enzymes, and is likely irrelevant to function. However, several systems including protein translocation, adenine nucleotide translocase, cytochrome c, and complex IV protein phosphorylation have been well correlated with enzymatic function along with the classical dehydrogenase systems. The second task is to review the current understanding of the kinase/phosphatase system within the matrix. Though it is clear that protein phosphorylation occurs within the matrix, based on 32 P incorporation and quantitative mass spectrometry measures, the kinase/phosphatase system responsible for this process is ill-defined. An argument is presented that remnants of the much more labile bacterial protein phosphoryl transfer system may be present in the matrix and that the evaluation of this possibility will require the application of approaches developed for bacterial cell signaling to the mitochondria.kinase; phosphatase; citric acid cycle; oxidative phosphorylation; protein transport; mitochondria intermembrane space; phosphohistidine; adenine nucleotide translocase; pyruvate dehydrogenase; branched chain ␣-ketoacid dehydrogenase THIS REVIEW ARTICLE is part of a collection on Post-translational Protein Modification in Metabolic Stress. Other articles appearing in this collection, as well as a full archive of all Review collections, can be found online at http://ajpheart. physiology.org/.

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Protein phosphorylation/dephosphorylation in the inner membrane of potato tuber mitochondria

Cited by 43 publications

References 20 publications

Identification of tyrosine-phosphorylated proteins of the mitochondrial oxidative phosphorylation machinery

Identification of tyrosine-phosphorylated proteins of the mitochondrial oxidative phosphorylation machinery

Phosphorylation of Formate Dehydrogenase in Potato Tuber Mitochondria

Cardiac mitochondrial matrix and respiratory complex protein phosphorylation

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