Purification and characterisation of a novel iso-propanol dehydrogenase from Phytomonas sp.

Uttaro, Antonio D.; Opperdoes, Frederik

doi:10.1016/s0166-6851(97)02830-2

Cited by 14 publications

(17 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…performed by checking the presence of marker enzymes (pyruvate kinase, hexokinase and alanine aminotransferase), according to Uttaro and Opperdoes (1997). The compartmentalization becomes significant if we accept that the main role of SOD is to detoxify endogenous superoxide anions and that the different biochemical processes that produce oxidizing agents inside the cell may need various antioxidant systems to protect macromolecules and organelles from oxidative damage.…”

Section: Discussionmentioning

confidence: 99%

“…Recovery (based on the sum of fractions) and relative specific activity (RSA) were calculated. For the correct identification of the subcellular fractions, we checked specific marker enzymes according to Uttaro and Opperdoes (1997), i.e., pyruvate kinase for the cytosol, hexokinase for the glycosome and alanine transferase for the mitochondria.…”

Section: Methodsmentioning

confidence: 99%

“…After incubation for 5 min at 4ºC, suspensions were centrifuged. Aliquots of the supernatants were used to assay the various enzymatic activities according to Uttaro and Opperdoes (1997).…”

Section: Methodsmentioning

confidence: 99%

“…Buffer 3 (50 mM potassium phosphate, pH 7.8) was chosen for these determinations. Pyruvate kinase, hexokinase and alanine aminotransferase activities were determined according to Uttaro and Opperdoes (1997).…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

Mateo

Marı́n

Pérez-Cordón

et al. 2008

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

Mateo

Marı́n

Pérez-Cordón

et al. 2008

Mem. Inst. Oswaldo Cruz

View full text Add to dashboard Cite

show abstract

“…It is a dimeric NAD-linked enzyme able to use a broad range of both primary and secondary alcohols. We have named the enzyme isopropyl alcohol dehydrogenase (iPDH) because of its high affinity and specific activity with this substrate (9). To understand the physiological role of the enzyme and its peculiar properties, we decided to clone the genes and to express the corresponding proteins in a heterologous system.…”

Section: Alcohol Dehydrogenases (Adh)mentioning

confidence: 99%

The Multifunctional Isopropyl Alcohol Dehydrogenase of Phytomonas sp. Could Be the Result of a Horizontal Gene Transfer from a Bacterium to the Trypanosomatid Lineage

Molinas

Altabe

Opperdoes

et al. 2003

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Isopropyl alcohol dehydrogenase (iPDH) is a dimeric mitochondrial alcohol dehydrogenase (ADH), so far detected within the Trypanosomatidae only in the genusPhytomonas. The cloning, sequencing, and heterologous expression of the two gene alleles of the enzyme revealed that it is a zinc-dependent medium-chain ADH. Both polypeptides have 361 amino acids. A mitochondrial targeting sequence was identified. The mature proteins each have 348 amino acids and a calculated molecular mass of 37 kDa. They differ only in one amino acid, which can explain the three isoenzymes and their respective isoelectric points previously found. A phylogenetic analysis locates iPDH within a cluster with fermentative ADHs from bacteria, sharing 74% similarity and 60% identity with Ralstonia eutropha ADH. The characterization of the two bacterially expressed Phytomonas enzymes and the comparison of their kinetic properties with those of the wild-type iPDH and of the R. eutropha ADH strongly support the idea of a horizontal gene transfer event from a bacterium to a trypanosomatid to explain the origin of the iPDH in Phytomonas. Phytomonas iPDH and R. eutropha ADH are able to use a wide range of substrates with similar K m values such as primary and secondary alcohols, diols, and aldehydes, as well as ketones such as acetone, diacetyl, and acetoin. We speculate that, as for R. eutropha ADH, Phytomonas iPDH acts as a safety valve for the release of excess reducing power. Alcohol dehydrogenases (ADH)1 display a wide range of substrate specificities and fulfil several key physiological functions. They are classified into three major categories: NAD(P)-dependent enzymes, NAD(P)-independent enzymes, and irreversible alcohol oxidases. NAD(P)-dependent ADHs are subdivided into groups according to their metal dependence: medium-chain zinc-dependent, short-chain zinc-independent, and iron-activated enzymes (1).Zinc-dependent medium-chain dehydrogenases constitute a large superfamily of enzymes widely found in vertebrates, plants, fungi, and bacteria, including polyol dehydrogenases, threonine dehydrogenase, quinone oxidoreductases, and other proteins (2). Currently, they can be divided into tetrameric (as NAD-dependent ADHs from fungi or some bacterial NADP-dependent enzymes) and dimeric NAD-linked ADH families. The last, in turn, are divided into the classes found in vertebrates (I-VII) or plants (III, C and P), according to substrate preference and amino acid identity (3, 4).Although it is not uncommon for ADHs to exhibit a broad substrate spectrum, as a rule they are specific either for primary or for secondary alcohols. One interesting exception is the tetrameric multifunctional Ralstonia eutropha (formerly Alcaligenes eutrophus) ADH. This enzyme is able to use both primary and secondary alcohols, polyols such as 2,3-butanediol, acetaldehyde, acetone, diacetyl, or acetoin as substrates and to use NAD(H) or NADP(H) as cosubstrate (5).Few trypanosomatid ADHs have been described. These are the cytosolic NADPH-linked aldehyde reductases from Leishmani...

show abstract

Rotenone-sensitive mitochondrial potential in Phytomonas serpens: electrophoretic Ca2+ accumulation

Moysés

Barrabin

2004

Biochimica et Biophysica Acta (BBA) - Bioenergetics

View full text Add to dashboard Cite

Phytomonas sp. are flagellated trypanosomatid plant parasites that cause diseases of economic importance in plantations of coffee, oil palm, cassava and coconuts. Here we investigated Ca(2+) uptake by the vanadate-insensitive compartments using permeabilized Phytomonas serpens promastigotes. This uptake occurs at a rate of 1.13+/-0.23 nmol Ca(2+) mg x protein(-1) min(-1). It is completely abolished by the H(+) ionophore FCCP and by valinomycin and nigericin. It is also inhibited by 2 microM ruthenium red, which, at this low concentration, is known to inhibit the mitochondrial calcium uniport. Furthermore, salicylhydroxamic acid (SHAM) and propylgallate, specific inhibitors of the alternative oxidase in plant and parasite mitochondria, are also effective as inhibitors of the Ca(2+) transport. These compounds abolish the membrane potential that is monitored with safranine O. Rotenone, an inhibitor of NADH-CoQ oxidoreductase, can also dissipate 100% of the membrane potential. It is suggested that the mitochondria of P. serpens can be energized via oxidation of NADH in a pathway involving the NADH-CoQ oxidoreductase and the alternative oxidase to regenerate the ubiquinone. The electrochemical H(+) gradient can be used to promote Ca(2+) uptake by the mitochondria.

show abstract

Purification and characterisation of a novel iso-propanol dehydrogenase from Phytomonas sp.

Cited by 14 publications

References 14 publications

Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

Purification and biochemical characterization of four iron superoxide dismutases in Trypanosoma cruzi

The Multifunctional Isopropyl Alcohol Dehydrogenase of Phytomonas sp. Could Be the Result of a Horizontal Gene Transfer from a Bacterium to the Trypanosomatid Lineage

Rotenone-sensitive mitochondrial potential in Phytomonas serpens: electrophoretic Ca2+ accumulation

Contact Info

Product

Resources

About