Purification, Characterization, and Submitochondrial Localization of the 32-Kilodalton NADH Dehydrogenase from Maize

Knudten, Andrew F.; Thelen, Jay J.; Luethy, Michael H.; Elthon, Thomas E.

doi:10.1104/pp.106.3.1115

Cited by 16 publications

(18 citation statements)

References 26 publications

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“…Recently, it has been suggested that in beetroot mitochondria external rotenone‐insensitive NAD(P)H oxidation is catalysed by either a 58 kDa and/or 32 kDa protein whilst matrix rotenone‐insensitive NADH oxidation occurs via a 43 kDa dimer [8–10]. Sonication studies have shown that both the matrix and external enzymes are peripherally bound to the inner‐membrane of plant mitochondria.…”

Section: Introductionmentioning

confidence: 99%

The nuclear origin of the non‐phosphorylating NADH dehydrogenases of plant mitochondria

et al. 1999

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The oxidation of matrix and cytosolic NADH by isolated beetroot and wheat leaf mitochondria was investigated to determine whether the rotenone-insensitive NADH dehydrogenases of plant mitochondria were the products of nuclear or mitochondrial genes. After aging beetroot tissue (slicing and incubating in a CaSO 4 solution), the induction of the level of matrix NADH oxidation in the presence of rotenone was greatly reduced in mitochondria isolated from tissue treated with cycloheximide, a nuclear protein synthesis inhibitor. This was also true for the oxidation of cytosolic NADH. Mitochondria isolated from chloramphenicol-treated tissue exhibited greatly increased levels of both matrix and external rotenone-insensitive NADH oxidation when compared to the increase due to the aging process alone. This increase was not accompanied by an increase in matrix NAD-linked substrate dehydrogenases such as malic enzyme nor intra-mitochondrial NAD levels. Possible explanations for this increase in rotenone-insensitive NADH oxidation are discussed. Based on these results we have concluded that the matrix facing rotenone-insensitive NADH dehydrogenase of plant mitochondria is encoded by a nuclear gene and synthesis of the protein occurs in the cytosol.z 1999 Federation of European Biochemical Societies.

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Section: Introductionmentioning

confidence: 99%

The nuclear origin of the non‐phosphorylating NADH dehydrogenases of plant mitochondria

et al. 1999

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“…Antibodies were prepared against the 58-kD protein from red beet roots and used in immunoprecipitation experiments under conditions as described by Knudten et al (1994). The antibodies were incubated with maize Mono Q fractions containing the 58-kD NAD(P)H DH activity and found to immunoprecipitate 44% of the activity.…”

Section: Immunological Characterization Of the 58-kd Nad(p)h Dhmentioning

confidence: 99%

“…In freshly isolated Procedures for cross-linking and for isolation of outer membranes were as described by Knudten et al (1994). MITOS, Mitochondria; X-LINKED MITOS, cross-linked mitochondria.…”

Section: Submitochondrial Localization Of the 58-kd Nad(p)h Dh From Mmentioning

confidence: 99%

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Purification, Characterization, and Submitochondrial Localization of a 58-Kilodalton NAD(P)H Dehydrogenase

et al. 1995

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An NADH dehydrogenase activity from red beet (Befa vulgaris 1.) root mitochondria was purified to a 58-kD protein doublet. An immunologically related dehydrogenase was partially purified from maize (Zea mays 1. 873) mitochondria to a 58-kD protein doublet, a 45-kD protein, and a few other less prevalent proteins. Polyclonal antibodies prepared against the 58-kD protein of red beet roots were found to immunoprecipitate the NAD(P)H dehydrogenase activity. The antibodies cross-reacted to similar proteins in mitochondria from a number of plant species but not to rat liver mitochondrial proteins. The polyclonal antibodies were used in conjunction with maize mitochondrial fractionation to show that the 58-kD protein was likely part of a protein complex loosely associated with the membrane fraction. A membrane-impermeable protein crosslinking agent was used to further show that the majority of the 58-kD protein was located on the outer surface of the inner mitochondrial membrane or in the intermembrane space. Analysis of the cross-linked 58-kD NAD(P)H dehydrogenase indicated that specific proteins of 64, 48, and 45 kD were cross-linked to the 58-kD protein doublet. The NAD(P)H dehydrogenase activity was not affected by ethyleneglycol-bis(P-aminoethyl ether)-N,N'-tetraacetic acid or CaCI,, was stimulated somewhat (21 %) by flavin mononucleotide, was inhibited by pchloromercuribenzoic acid (49%) and mersalyl (40%), and was inhibited by a bud scale extract of Platanus occidentalis L. containhg plataneth (61 %).In contrast to their mammalian counterparts, plant mitochondria are capable of oxidizing cytoplasmic NAD(P)H directly, coupling this oxidation to the electron transport chain (Moller and Lin, 1986). This occurs via exogenous NAD(P)H DHs located on the cytosolic face of the inner mitochondrial membrane (Palmer and Moller, 1982;Moller, 1986;Moller and Lin, 1986; Douce and Neuberger, 1989). The oxidation of endogenous mitochondrial matrix substrates (NADH and succinate) has been shown to take precedence over the oxidation of exogenous NAD(P)H (Dry et al., 1983;Day et al., 1985). Thus, it has been suggested that the exogenous NAD(P)H DHs may function in balancing the redox levels between NAD(P)H pools in the cytosol and the mitochondrion (Moller and Lin, 1986; Douce and Neuberger, 1989). As a result, the exogenous NAD(P)H DH could coordinate glycolytic flwc with flow through the Krebs cycle. If this is true, then regulation of exogenous NAD(P)H DH activity could greatly influence plant metabolism. Consistent with this hypothesis, Kromer and Heldt (1991) have shown that the oxidation by mitochondria of reducing equivalents generated during photosynthesis is vital for obtaining maximum photosynthetic rates. Exogenous NAD(P)H DH activity can be distinguished from the other mitochondrial NAD(P)H DH activities by its insensitivity to rotenone (Wilson and Hanson, 1969), stimulation by Ca2' and inhibition by EGTA (Coleman and Palmer, 1971), and sensitivity to platanetin (Ravanel et al., 1986). Although these characteristics ca...

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“…Furthermore, it is not known whether all of these enzymes exist in Arabidopsis. A number of attempts at purification of these proteins have provided little consensus as to the size and number of polypeptides involved (Klein and Burke, 1984;Luethy et al, 1991;Rasmusson et al, 1993;Knudten et al, 1994;Day, 1996a, 1996b).…”

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confidence: 99%

Identification of AtNDI1, an Internal Non-Phosphorylating NAD(P)H Dehydrogenase in Arabidopsis Mitochondria

et al. 2003

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Plant mitochondria contain non-phosphorylating NAD(P)H dehydrogenases (DHs) that are not found in animal mitochondria. The physiological function, substrate specificity, and location of enzymes within this family have yet to be conclusively determined. We have linked genome sequence information to protein and biochemical data to identify that At1g07180 (SwissProt Q8GWA1) from the Arabidopsis Genome Initiative database encodes AtNDI1, an internal NAD(P)H DH in Arabidopsis mitochondria. Three lines of evidence are presented: (a) The predicted protein sequence of AtNDI1 has high homology with other designated NAD(P)H DHs from microorganisms, (b) the capacity for matrix NAD(P)H oxidation via the rotenone-insensitive pathway is significantly reduced in the Atndi1 mutant plant line, and (c) the in vitro translation product of AtNDI1 is imported into isolated mitochondria and located on the inside of the inner membrane.

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Purification, Characterization, and Submitochondrial Localization of the 32-Kilodalton NADH Dehydrogenase from Maize

Cited by 16 publications

References 26 publications

The nuclear origin of the non‐phosphorylating NADH dehydrogenases of plant mitochondria

The nuclear origin of the non‐phosphorylating NADH dehydrogenases of plant mitochondria

Purification, Characterization, and Submitochondrial Localization of a 58-Kilodalton NAD(P)H Dehydrogenase

Identification of AtNDI1, an Internal Non-Phosphorylating NAD(P)H Dehydrogenase in Arabidopsis Mitochondria

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