Trypanosoma brucei: Preparation and some properties of a multienzyme complex catalysing part of the glycolytic pathway

Oduro, K.K.; Bowman, I.B.R.; Flynn, I.W.

doi:10.1016/0014-4894(80)90025-9

Cited by 42 publications

(18 citation statements)

References 11 publications

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“…The fact that PGK's localization is either glycosomal or cytosolic – or both, as observed in promastigotes of different Leishmania species, T. cruzi epimastigotes and T. borreli [36-39] – as required to maintain the glycosomal ATP/ADP balance, offers a rationale for finding phosphoglycerate mutase and enolase always in the cytosol [5,40-42]. Although the localization of these enzymes is not directly relevant for the ATP/ADP and NAD + /NADH balances, it would not provide any selective advantage to have them inside the glycosomes with a variable compartmentation of PGK and a constitutive expression of PYK in the cytosol.…”

Section: Metabolic Peculiarities Which Are Shared By All Kinetoplastimentioning

confidence: 99%

“…The evolutionary origin of glycosomes has been a matter of interest since the identification of these organelles, over 25 years ago [5,40,77]. Our original hypothesis proposed that the glycosome was derived from a bacterial endosymbiont, in a manner similar to mitochondria and chloroplasts, but that it had lost its entire genome [78].…”

Section: How Did Glycosomes Originate?mentioning

confidence: 99%

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Evolution of energy metabolism and its compartmentation in Kinetoplastida

Hannaert

Bringaud

Opperdoes

et al. 2003

Kinetoplastid Biol Dis

154

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Kinetoplastida are protozoan organisms that probably diverged early in evolution from other eukaryotes. They are characterized by a number of unique features with respect to their energy and carbohydrate metabolism. These organisms possess peculiar peroxisomes, called glycosomes, which play a central role in this metabolism; the organelles harbour enzymes of several catabolic and anabolic routes, including major parts of the glycolytic and pentosephosphate pathways. The kinetoplastid mitochondrion is also unusual with regard to both its structural and functional properties.In this review, we describe the unique compartmentation of metabolism in Kinetoplastida and the metabolic properties resulting from this compartmentation. We discuss the evidence for our recently proposed hypothesis that a common ancestor of Kinetoplastida and Euglenida acquired a photosynthetic alga as an endosymbiont, contrary to the earlier notion that this event occurred at a later stage of evolution, in the Euglenida lineage alone. The endosymbiont was subsequently lost from the kinetoplastid lineage but, during that process, some of its pathways of energy and carbohydrate metabolism were sequestered in the kinetoplastid peroxisomes, which consequently became glycosomes. The evolution of the kinetoplastid glycosomes and the possible selective advantages of these organelles for Kinetoplastida are discussed. We propose that the possession of glycosomes provided metabolic flexibility that has been important for the organisms to adapt easily to changing environmental conditions. It is likely that metabolic flexibility has been an important selective advantage for many kinetoplastid species during their evolution into the highly successful parasites today found in many divergent taxonomic groups.Also addressed is the evolution of the kinetoplastid mitochondrion, from a supposedly pluripotent organelle, attributed to a single endosymbiotic event that resulted in all mitochondria and hydrogenosomes of extant eukaryotes. Furthermore, indications are presented that Kinetoplastida may have acquired other enzymes of energy and carbohydrate metabolism by various lateral gene transfer events different from those that involved the algal-and α-proteobacterial-like endosymbionts responsible for the respective formation of the glycosomes and mitochondria.

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Section: Metabolic Peculiarities Which Are Shared By All Kinetoplastimentioning

confidence: 99%

Section: How Did Glycosomes Originate?mentioning

confidence: 99%

Evolution of energy metabolism and its compartmentation in Kinetoplastida

Hannaert

Bringaud

Opperdoes

et al. 2003

Kinetoplastid Biol Dis

154

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“…Such an organisation would lead to a more efficient catalysis of glycolysis than if the glycosome simply consisted of a bag filled with a soup of enzymes and intermediates. One possible organisation could be a multienzyme complex consisting of all nine enzymes in a ratio of their abundancy and some evidence has been presented for the existence of such complexes [8,9]. With the quantitative data now available we have tried to make an estimate of the composition and the size of such a complex.…”

Section: Concentration and Organization Of Glycosomal Constituentsmentioning

confidence: 99%

Glycolytic enzymes of Trypanosoma brucei. Simultaneous purification, intraglycosomal concentrations and physical properties

1986

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We have developed a method for the simultaneous purification of hexokinase, glucosephosphate isomerase, phosphofructokinase, fructose-l,6-bisphosphate aldolase, triosephosphate isomerase, D-glyceraldehyde-phosphate dehydrogenase, phosphoglycerate kinase, glycerol-3-phosphate dehydrogenase and glycerol kinase from Trypanosoma brucei in yields varying over 8 -55%.Crude glycosomes were prepared by differential centrifugation of cell homogenates. Subsequent hydrophobic interaction chromatography on phenyl-Sepharose resulted in six pools containing various mixtures of enzymes. These pools were processed via affinity chromatography (immobilized ATP), hydrophobic interaction chromatography (octyl-Sepharose) and ion-exchange chromatography (CM-and DEAE-cellulose) which resulted in the purification of all nine enzymes.The native enzyme and subunit molecular masses, as determined by gel filtration and gel electrophoresis under denaturing conditions, were compared with those of their homologous counterparts from other organisms. Trypanosomal hexokinase is a hexamer and differs in subunit composition from the mammalian enzymes (monomers) as well as in subunit size (51 kDa versus 96-100 kDa, respectively). Phosphofructokinase only differs in subunit size (51 kDa for T. brucei versus 80 -90 kDa for mammals) but had identical subunit composition (tetrameric). The others all have the same subunit composition as their mammalian counterparts. Except for triosephosphate isomerase, all Trypanosoma enzymes have subunits which are 1 -5 kDa larger in size.Together these nine enzymes contribute 3.3 f 1.6% to the total cellular protein of T. brucei and at least 90% to the total glycosomal protein. A comparison of calculated intraglycosomal concentrations of the enzymes with the glycosomal metabolite concentrations shows that in the case of aldolase, glyceraldehyde-phosphate dehydrogenase and phosphoglycerate kinase, the concentration of active sites is of the same order of magnitude as that of their reactants.A common feature of the glycosomal glycolytic enzymes (with the exception of glucosephosphate isomerase) is that they are highly basic proteins with PI values between 8.8 and 10.2, values which are 1-4 higher than in the case of their mammalian cytosolic counterparts and 3 -6 higher than in the case of the various unicellular organisms.It is suggested that both the larger subunit size and the basic character of the T. brucei glycolytic proteins are involved in the routing of the enzymes from their site of biogenesis (the cytosol) towards their site of action (the glycosome).

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“…Glycosomal enzymes of T. brucei differ from other glycolytic enzymes in that they have a high tendency to form multi-enzyme complexes. Such complexes are stable and can easily be isolated in the absence of a surrounding membrane (Opperdoes and Nwagwu, 1980;Oduro et al, 1980;Aman et al, 1985). Therefore, at least some of the elements unique to the glycosomal enzymes might be involved in protein -protein interactions of the constituent enzymes of such complexes.…”

Section: Net Charges Of Glycosomal Enzymesmentioning

confidence: 99%

Common elements on the surface of glycolytic enzymes from Trypanosoma brucei may serve as topogenic signals for import into glycosomes.

Wierenga¹,

Swinkels²,

Michels³

et al. 1987

The EMBO Journal

105

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In Trypanosoma brucei, a major pathogenic protozoan parasite of Central Africa, a number of glycolytic enzymes present in the cytosol of other organisms are uniquely segregated in a microbody-like organelle, the glycosome, which they are believed to reach post-translationally after being synthesized by free ribosomes in the cytosol. In a search for possible topogenic signals responsible for import into glycosomes we have compared the amino acid sequences of four glycosomal enzymes: triosephosphate isomerase (TIM), glyceraldehyde-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK) and aldolase (ALDO), with each other and with their cytosolic counterparts. Each of these enzymes contains a marked excess of positive charges, distributed in two or more clusters along the polypeptide chain. Modelling of the three-dimensional structures of TIM, PGK and GAPDH using the known structural coordinates of homologous enzymes from other organisms indicates that all three may have in common two 'hot spots' about 40 A apart, which themselves include a pair of basic amino acid residues separated by a distance of about 7 A. The sequence of glycosomal ALDO, for which no three-dimensional information is available, is compatible with the presence of the same configuration on the surface of this enzyme. We propose that this feature plays an essential role in the import of enzymes into glycosomes.

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Trypanosoma brucei: Preparation and some properties of a multienzyme complex catalysing part of the glycolytic pathway

Cited by 42 publications

References 11 publications

Evolution of energy metabolism and its compartmentation in Kinetoplastida

Evolution of energy metabolism and its compartmentation in Kinetoplastida

Glycolytic enzymes of Trypanosoma brucei. Simultaneous purification, intraglycosomal concentrations and physical properties

Common elements on the surface of glycolytic enzymes from Trypanosoma brucei may serve as topogenic signals for import into glycosomes.

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