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Regulation of the mitochondrial tricarboxylic acid cycle

Adriano Nunes-Nesi1,2, Wagner L Araujo1, Toshihiro Obata3 and
Alisdair R Fernie3

Recent years have seen considerable advances in our cells that were first described by Hans Krebs in pigeon
understanding of the particular physiological roles of the muscle [5], relatively little is known concerning the
constituent enzymes of the tricarboxylic acid (TCA) cycle. regulation and control of this pathway [1,6!,7!].
Despite acquiring a fairly comprehensive overview of the
functional importance of these proteins relatively little is known The tricarboxylic acid (TCA) cycle is composed by a set of
concerning how this important pathway is regulated. In this eight enzymes primarily linking the product of the oxi-
review we concentrate on the mitochondrial reactions since dation of pyruvate and malate (generated in the cytosol) to
this organelle is the only one in which a full cycle can, at least CO2 with the generation of NADH for the oxidation by the
theoretically, operate. We summarize what is known about the mitochondrial respiratory chain [8]. The genomic organ-
regulation of the enzymes of the pathway both from historical ization and the subcellular localization of the enzymes
kinetic studies as well as discussing more recent transcriptional involved in the Arabidopsis TCA cycle are summarized
and proteomic studies and our enhanced understanding of in Supplementary Table 1; however those are not
subcellular compartmentation within the context of metabolic described in detail since firstly it has been expertly
regulation. reviewed elsewhere [9] and secondly we intend to con-
Addresses centrate here on the mitochondrial reactions since this
Departamento de Biologia Vegetal, Universidade Federal de Vicosa, organelle is the only one in which a full cycle can, at least
36570-000 Vicosa, Minas Gerais, Brazil
theoretically, operate [10!]. Whilst the presence of organic
Max-Planck Partner Group, Departamento de Biologia Vegetal,
acids, particularly TCA cycle intermediates, in all plants is
Universidade Federal de Vicosa, 36570-000 Vicosa, Minas Gerais, Brazil
Max-Planck-Institut fur Molekulare Pflanzenphysiologie, Am known to support numerous and diverse functions within
Muhlenberg 1, 14476 Potsdam-Golm, Germany and beyond cellular metabolism, the level of accumulation
of the various organic acids is extremely variable between
Corresponding author: Fernie, species, developmental stages and tissue types [11] pro-
Alisdair R (
viding further support that the enzymes involved in the
interconversion of these metabolic intermediates are sub-
Current Opinion in Plant Biology 2013, 16:335343 ject to tight regulatory control. The sequestration of
This review comes from a themed issue on Physiology and
organic acids into the vacuole and secretion into the rhizo-
metabolism sphere additionally represent important mechanisms of
Edited by John Browse and Edward Farmer
regulating these intermediates; however, these have been
recently reviewed elsewhere [11,12].
For a complete overview see the Issue and the Editorial
Available online 22nd February 2013
Hints to the regulation of the TCA cycle have been
1369-5266/$ see front matter, # 2013 Elsevier Ltd. All rights provided by a recent metabolic control analysis which
show that much of the control through this pathway is resident in fumarase, malate dehydrogenase (MDH) and
2-oxoglutarate dehydrogenase [1], suggesting that these
would be sensitive targets for flux regulation. A lack of
Introduction subcellular information concerning the levels of inter-
There is now considerable cumulative evidence that mediates of the cycle [13] currently precludes us from
mitochondrial respiratory function is associated with being able to assess the potential of the constituent
proper maintenance of cellular metabolism as a whole. enzymes to play regulatory roles on the basis of disequi-
Reverse molecular genetic approaches have proven librium ratios. However, there is a growing body of
particularly instrumental in establishing the physiological information concerning allosteric and post-translational
and metabolic basis for mitochondrial function in living regulation of specific enzymes from which mechanisms of
plant cells [1]. This was particularly true for increasing our regulation can be inferred (Figure 1). We shall discuss
knowledge of mitochondrial metabolism as of fundamen- these studies in two separate sections. In the first we will
tal importance in many other important cellular processes describe focused in vitro and in vivo studies investigating
such as photosynthesis, photorespiration, nitrogen metab- the role of enzyme effectors. The second section will
olism, redox regulation and signalling [24]. That said, focus on broad-based studies in which changes in tran-
and quite remarkably since it was demonstrated over 50 scripts, protein abundance, post-translational modifi-
years ago that exactly the same reactions occur in plant cation of proteins or metabolite levels were assessed in Current Opinion in Plant Biology 2013, 16:335343

336 Physiology and metabolism

Figure 1

1 Pyruvate 2 Phosphorylation
Redox regulation
TCA cycle
cycle Isocitrate

Highly sensitive to
oxidative stress and NO

3 Phosphorylation 4 Phosphorylation
Redox regulation Redox regulation

cycle Isocitrate cycle
2-OG light dependent
2-OG SucCoA
2-OGDH regulation

5 6
Phosphorylation Phosphoryation
Redox regulation Redox regulation
cycle cycle
Succinate Succinate
light dependent Phytochrome and
SucCoA regulation nitric oxid regulation

7 8 OAA Phosphorylation
Malate MDH Redox regulation
FUM Malate

Fumarate TCA TCA

cycle cycle

Current Opinion in Plant Biology

Current known regulatory mechanisms of the mitochondrial TCA cycle. Each enzymatic step of the TCA cycle is represented having only the enzyme
and its substrate and product whilst specific non-metabolite-mediated regulatory mechanisms are specified. Mechanisms involved in the
transcriptional regulation level for each enzyme are demonstrated in light blue boxes whilst post-transcriptional mechanism are shown in black boxes.
For clarity, cofactors have been omitted the 2-OGDH reaction requires CoA; the CS and SCoA L reactions produce CoA; NAD+ is converted into
NADH by the IDH, 2-OGDH and MDH reactions; ADP is converted into ATP by the SCoA L reaction. CS, citrate synthase (note this actually catalyses
the condensation of acetyl CoA and OAA; pyruvate is just shown to indicate its entrance point); ACO, aconitase; IDH, isocitrate dehydrogenase; 2-
OGDH, 2-oxoglutarate dehydrogenase complex; SCoAL, succinyl CoA ligase; SDH, succinate dehydrogenase; FUM, fumarase; MDH, malate
dehydrogenase; OAA, oxaloacetate; 2-OG, 2-oxoglutarate; TCA cycle, tricarboxylic acid cycle.

response to environmental or experimental perturbation TCA cycle effectors

and from which potential elements of regulation were In the following paragraphs we will briefly detail the
identified. Finally we discuss regulatory principles which known metabolite regulators of each enzyme which are
have already been described in non-plant systems and additionally summarized as an overview in Table 1. For
conclude with a perspective for future efforts aimed at the purpose of the discussion, we have also included the
unraveling the regulation of this vital pathway. pyruvate dehydrogenase complex (PDC) reaction which

Current Opinion in Plant Biology 2013, 16:335343

Regulation of the mitochondrial tricarboxylic acid cycle Nunes-Nesi et al. 337

Table 1

Summary of known metabolite-mediated regulatory characteristics of the TCA cycle enzymes

Enzyme of the pathway Effectors Inhibitors

Pyruvate dehydrogenase Pyruvate, thiamine pyrophosphate NADH, acetyl-CoA, NH4, glyoxylate
Citrate synthase ADP ATP, NADH and succinyl-CoA
Aconitase n.d. Citramalate, oxalomalate, NADH, H2O2
Isocitrate dehydrogenase ADP, NAD+ and Ca2+ ATP and NAD(P)H, succinic semialdehyde, succinate,
glutamate, GABA
2-Oxoglutarate dehydrogenase Thiamine pyrophosphate, AMP, NADH
ADP; NAD+ and Ca2+
Succinyl-CoA ligase 2-Oxoglutarate NADH. Malonate, citrate, isocitrate
Succinate dehydrogenase ATP, ADP, NADH, FAD, QH2 Oxaloacetate; NADH, nitric oxide
Fumarase n.d. ATP, ADP and AMP; NADH, pyruvate, citrate, 2-oxoglutarate
Malate dehydrogenase n.d. NADH

n.d.: not determined; GABA: g-aminobutyric acid.

is not strictly part of the TCA cycle but is, nevertheless, enzymes as well as by the PDC, it is reasonable to assume
intimately associated with it. The PDC is a complex that mitochondrial NADH/NAD+ ratio has a major impact
consisting of three components: pyruvate dehydrogenase on the flux through the TCA cycle. However, it is
(E1), dihydrolipoyl acetyltransferase (E2) and dihydroli- important to note both for this enzyme and the other
poyl dehydrogenase (E3). There are two distinct and dehydrogenases of the TCA cycle (discussed below) that
spatially separated forms of the PDC in plant cells; elegant studies from the Mller laboratory have revealed
mitochondrial PDC and plastidial PDC. The mitochon- that the free NADH concentration is kept constant in
drial form of PDC is one of the carbon entry sites into the plant mitochondria under different metabolic conditions
TCA cycle, whilst the plastidial PDC provides acetyl- rendering it important to interpret implications of in vitro
CoA and NADH for de novo fatty acid biosynthesis [14]. kinetics with caution [18]. In addition to this regulation
Moreover, the mitochondrial PDC has two associated the mitochondrial PDC is also regulated by product
regulatory enzymes: pyruvate dehydrogenase kinase inhibition by acetyl CoA [14] and activated by thiamine
(PDK) and phospho-pyruvate dehydrogenase phospha- pyrophosphate [19!!].
tase [14]. Given the exquisite regulation of this enzyme it
is generally assumed to be a major regulatory point of flux Citrate synthase (CS) is confined to the mitochondrial
through the cycle; however, as the discussion below matrix, except in tissues converting fatty acids into sugars
indicates it is certainly not the only regulatory point in which a peroxisomal isoform is also functional [20].
and may not even be the major one. Intriguingly, down Interestingly, this peroxisomal isoform is up-regulated
regulation of the inhibitory PDK in Arabidopsis resulted following the antisense repression of the mitochondrial
in plants displaying altered vegetative growth with isoform in tomato, and therefore able to functionally
reduced accumulation of vegetative tissues, early flower compensate the absence of the mitochondrial one [21].
development and shorter generation time [15], whilst a Structural and expression characteristics of mitochondrial
mutation of the E2 subunit, conferring an opposite effect isoforms from diverse species have been studied and the
on PDC activity, also resulted in a decreased organ growth identification of potential inter-domain disulfides in
[16]. The apparent contradictory nature of these obser- higher plant mitochondrial CS suggests paradoxical
vations suggests that further research will be needed to differences in redox-sensitivity relative to its animal
fully understand the physiological importance of the counterpart [22]. CS is also regulated by the cellular
phosphorylation of the PDC. What is less contentious NADH/NAD+ ratio, the ATP/ADP ratio and succinyl-
is the fact that the PDC activity interconnects glycolysis, CoA levels whilst being inhibited by ATP, NADH and
gluconeogenesis and fatty acid synthesis to the TCA succinyl-CoA and activated by ADP [23], suggesting that
cycle and is highly regulated by a range of allosteric its activity is tightly regulated at the metabolite level.
effectors [17]. It has long been demonstrated that PDC
as well as TCA cycle dehydrogenases displays product The next step of the TCA cycle is catalyzed by aconitase
inhibition in vitro by NADH [14]. Accordingly given that which catalyzes the reversible hydration of cis-aconitate to
the in vivo activities of PDC and other TCA cycle either citrate or isocitrate. In plants, aconitase has been
enzymes are responsive to the NADH/NAD+ ratio, this characterized in several tissues [24,25]. It has been also
provides a very sensitive mechanism by which it is demonstrated that at least in Arabidopsis two isoforms are
possible to tune balance the rate of pyruvate oxidation present in the mitochondria and one in the cytosol [26!].
by PDC and the TCA cycle activity with the rate of Interestingly whilst the number of genes encoding aco-
oxidative phosphorylation [17]. Because of the fact that nitase varies between plant species, the gene products are
NAD+ is a common co-factor utilized by three TCA cycle often dual targeted to both the mitochondria and the Current Opinion in Plant Biology 2013, 16:335343

338 Physiology and metabolism

cytosol [25,26!]. Notably the aconitase enzyme is highly pathway of porphyrin biosynthesis as well as being
sensitive to oxidative stress [27] since it has highly competitively inhibited by malonate in the reverse
oxidation sensitive iron (Fe)sulphur (S) cluster in its direction but is activated by 2-oxoglutarate and inhib-
active centre. Studies on citrus fruit vesicles and calli ited by both citrate as well as isocitrate and all down-
suggest that Fe availability may also be involved in the stream intermediates of the TCA cycle when assayed in
regulation of cytosolic aconitase activity; however the the forward direction [37]. Taken together these data
mitochondrial isoform is not affected by Fe limitation, thus suggest that the reaction catalyzed by ScoAL is
suggesting a distinct response of different aconitase likely co-ordinately regulated with those catalyzed by
forms to Fe shortage [28]. It has furthermore been other enzymes of the TCA cycle.
demonstrated that the lack of manganese superoxide
dismutase results in the inhibition of aconitase and the Succinate dehydrogenase (SDH), also commonly referred
subsequent enzyme of the cycle, isocitrate dehydrogen- as complex II, plays a key role in mitochondrial metab-
ase (IDH) [29]. Furthermore, the widely reported plant olism both as a member of the electron transport chain
metabolite citramalate can act as an endogenous inhibitor and TCA cycle [38]. The regulation of the enzyme was
of mitochondrial aconitase [30]. investigated in coupled mitochondria by simultaneously
measuring oxygen uptake rates and ubiquinone reduction
Isocitrate can subsequently be oxidatively decarboxy- levels [39]. This study revealed that the activation state
lated to 2-oxoglutarate by either NAD+-dependent or level of the enzyme is unambiguously reflected in the
NADP+-dependent IDH, generating CO2 and NADH or kinetic dependence of the succinate oxidation rate upon
NADPH, respectively [31]. In higher plants NADP+- the ubiquinone redox poise. Kinetic results indicated that
dependent IDH is located to the mitochondria, plastids, it is additionally activated by both ATP and ADP [39].
peroxisomes and cytosol [32], whereas NAD+-dependent Surprisingly NO, which has been described as an inhibi-
IDH is strictly mitochondrial and has been typically tor of plant respiratory chains owing to its high affinity for
associated with TCA cycle flux [33]. The enzyme iso- cytochrome c oxidase was recently demonstrated to act
lated from pea leaf mitochondria demonstrates product additionally on SDH [40].
inhibition by NADH, and is also inhibited by NADPH
[33]. In addition it has been observed that the latter The allosteric properties of fumarase from pea (Pisum
inhibition is non-competitive with NAD+ or NADH, sativum L.), revealed inhibition of this enzyme by phys-
suggesting that NADPH may be an allosteric regulator iological concentrations of pyruvate, 2-oxoglutarate and
since its Ki lies within the physiological range of mito- the adenine nucleotides ATP, ADP and AMP, consistent
chondrial NADPH concentrations [34]. In contrast to the with this step being an important control point of the
enzyme from non-plant sources plant NAD+-dependent TCA cycle [41]. Accordingly, down regulation of this
IDH is, however, unaffected by adenylates [33]. IDH is enzyme in tomato resulted in a relatively large reduction
believed to be an important regulatory step of the TCA in the rate of respiration in comparison to the majority of
cycle and plays an important role in maintaining the 2- other enzymes of the cycle [42].
oxoglutarate level and therefore in the regulation of
nitrogen assimilation [35]. The cycle is completed by MDH which catalyzes the
reversible oxidation of malate to produce oxaloacetate
The reaction catalyzed by the 2-oxoglutarate dehydro- (OAA) [43]. Whilst the equilibrium position favors malate
genase complex (OGDHC) represents a metabolic and NAD+ production, the in vivo removal of OAA by CS,
branch point connecting 2-oxoglutarate (and the TCA coupled with the removal of NADH by the respiratory
cycle) with nitrogen assimilation with 2-oxoglutarate chain, causes the reaction to function in the direction of
either being irreversibly degraded by the OGDHC or malate oxidation in most tissues [44]. Therefore, it is
providing carbon skeletons for nitrogen assimilation. likely that accumulation of NADH would lead to an
OGDHC is allosterically regulated in responses to second inhibition of the mitochondrial MDH.
messengers and metabolic indicators, such as Ca2+, ATP/
ADP, NADH/NAD+ and thiamine pyrophosphate [36]. To summarize the metabolic regulation of the TCA cycle
This complex is very similar to the PDC in the context of is somewhat complex and enzyme specific although sev-
its protein organization, cofactor dependency, and mech- eral of the enzymes are intrinsically coupled with the
anism of action. Although the OGDHC activity is not redox environment and the NADH/NAD+ ratio. Notably
known to be subject to covalent modification, its meta- the extent to which any metabolite mediated regulation
bolic regulation is quite complex, likely being regulated of TCA cycle flux is relevant in vivo is dependent on the
by energy charge, the NADH/NAD+ ratio, as well as by local concentrations of these metabolites. Given this fact
the levels of both its substrates and products [36]. it seems likely that further development of method-
ologies to determine subcellular metabolite concen-
The subsequent enzyme of the cycle, succinyl-CoA ligase trations [45] as well as a better understanding of the
(ScoAL), is feedback inhibited by intermediates of the mitochondrial carrier family of metabolite transporters

Current Opinion in Plant Biology 2013, 16:335343

Regulation of the mitochondrial tricarboxylic acid cycle Nunes-Nesi et al. 339

[46] and indeed transporters of the outer mitochondrial activity by phosphorylation of E1 subunit has been
membrane [47!!] will be crucial facilitators of our com- reported to be involved in the diurnal regulation of this
prehension of the regulation of flux through the pathway. enzyme complex. In Escherichia coli phosphorylation of
IDH is also known to play an important role in pathway
Mechanisms of regulation revealed by regulation [58]. In addition, large scale phosphoproteomic
profiling studies experiments revealed that many other TCA cycle
By contrast to the directed studies described above, we enzymes were also phosphorylated in Arabidopsis (e.g.
will now approach insights achieved from the application aconitase, IDH, OGDHC, ScoAL, SDH, and a mitochon-
of broad profiling techniques in a more general manner drial MDH; searched in PhosPhAt 4.0 database [59]). The
describing studies based at the transcript, protein and results thus render phosphorylation as a candidate mech-
finally metabolite level. At the transcriptional level it has anism for the TCA cycle regulation although the func-
been frequently observed that genes associated with the tionality of these phosphorylation events for in vivo
TCA cycle are down regulated during the night in an enzyme regulation remains to be tested.
essentially co-ordinated manner. However, a more recent
analysis [48] reveals that genes associated to the TCA The regulation of photosynthetic enzymes by the
cycle and mitochondrial electron transport chain actually reduction of regulatory disulfide bonds via ferredoxin
can be separated into three subgroups depending on their (Fdx)/thioredoxin (TRX) system has been well documen-
response to stress. A complete analysis of the expression ted in the plastid; however, plants additionally possess the
changes documented in public microarray databases at extraplastidic NADP/TRX system whereby NADPH
the level of tissue expression and response to stress is reduces TRX by means of the flavoenzyme NADP/
additionally provided in Supplementary Figures 1 and 2, TRX reductase (NTR) [60]. By contrast to the extensive
respectively. Owing to space limitations we will not studies on the regulatory role of TRX systems in the
comment on all of these suffice to say that there are plastid and the cytosol, we have scant understanding of
significant patterns of similarities and differences across the role of TRX in plant mitochondria. That said, pro-
both tissue type and the application of stress. Of particular teomic analyses of TRX binding proteins in Arabidopsis
note is the very high up-regulation of the TCA cycle mitochondria detected seven TCA cycle related enzymes
during osmotic, salt and UV-B stress consistent with [61!!], with other proteomic studies on identifying both
knowledge acquired in other experiments [4951]. Not cytosolic and mitochondrial MDH as targets of TRX [62].
visible in these datasets were the above mentioned These results suggest a direct redox regulation of TCA
responses to light; however, preliminary evidence cycle enzymes via NADP/TRX system as one of the
suggests that at least in the case of SDH differential regulatory mechanisms of this pathway. However, further
expression is regulated by the phytochrome pathway [52]. studies will be required in order to experimentally assess
the impact of redox regulation on the flux through the
The use of proteomics in understanding the TCA cycle TCA cycle. Lys acetylation of proteins has recently been
regulation has been two-fold. Firstly, diagnosing changes demonstrated to play an extensive role in the regulation
in protein abundance and secondly in the identification of metabolic enzymes [63].
and interpretation of post-translational modifications of
the proteins. Interestingly whilst there are several studies Proteomic studies revealed that all enzymes of TCA cycle
diagnosing the changes in mitochondrial proteome under are acetylated in human liver tissue and E. coli and the
stress conditions (reviewed in [53,54]) relatively few levels such as glucose, amino acids and fatty acids, gener-
changes in abundance of TCA cycle enzymes have been ally influence the acetylation status of metabolic enzymes
observed. Namely a decrease of fumarase and accumu- [64,65]. By contrast proteomic analysis of Arabidopsis
lation of a mitochondrial MDH and a subunit of PDC detected only a singly acetylated enzyme related to
under oxidative stress [55] and the increase of aconitase, TCA cycle, NAD+ dependent MDH which is localized
IDH, OGDHC E2 subunit and mitochondrial MDH in the cytosol [66]. Further effort should be made here
under flooding stress [56] were reported. On the other since as the authors state this most likely merely reflects
hand, abundance of TCA cycle enzymes showed dynamic that a lower depth of coverage of the Lys-acetylated
diurnal changes peaking early in the night, indicating four proteome was achieved in this study, rather than a lack
diurnal phases of the TCA cycle operation governed by of physiologically relevant Lys acetylation [66].
variation in the capacity of respiratory metabolism [57].
Interrogation of a recent compilation of metabolomics
Turning to post-translational modifications a huge num- datasets of plants exposed to abiotic stress reveals that the
ber of potential modifications of proteins are possible; cellular level of TCA cycle intermediates respond
however, we will limit our discussion here to phosphoryl- dramatically to several stresses [67]. Of particular note
ation, redox modification and protein acetylation since here given changes described above are the changes
these are arguably the best understood in terms of the following manipulation of the redox status [68], tissue
mitochondrial TCA cycle. Down regulation of PDC oxygenation level [69] and of elements of the circadian Current Opinion in Plant Biology 2013, 16:335343

340 Physiology and metabolism

clock [19!!,70]. Following the manipulation of the thiol- studies have indicated differential association of fumarase
disulphide status of Arabidopsis leaves by treatment with within protein complexes under conditions of oxidative
DTT there was a decrease in the metabolites aconitate, stress [78]. It seems likely that targeted approaches
isocitrate and 2-oxoglutarate but conversely an increase in designed specifically at the TCA cycle will be required
levels of succinate, fumarate and malate [68], consistent to better investigate the possibility that the plant cycle like
with the proteomic suggestion of direct redox regulation of its mammalian and bacterial counterparts also forms
the cycle. Similarly, metabolite profiling of waterlogging metabolite channels. To summarize, current understand-
induced hypoxia in Lotus japonica revealed the operation of ing of the regulation of the TCA cycle is that it is the result
a bifurcated TCA cycle pathway and an inhibition of the of an intricate interplay of several levels of regulation
reaction catalyzed by SDH [69]. Metabolomics on the including transcription, protein stability, protein modifi-
pseudo response regulator prr9 prr7 prr5 triple Ara- cations and allosteric effectors. Proteomics approaches
bidopsis mutants [70] and thiamin-pyrophosphate ribos- have clearly suggested that direct redox modulation of
witch deficient Arabidopsis [19!!] additionally provide several of the enzymes of the TCA cycle may well
strong support for the involvement of machinery of the represent a novel mechanism of regulation. It can be
circadian clock playing a role in the diurnal regulation of anticipated that further such mechanisms will be uncov-
the TCA cycle. In the latter case thiamine biosynthesis was ered as proteomics technologies mature. Two other devel-
demonstrated to be under the control of the circadian clock opments that we anticipate having great promise here are
and rather surprisingly that the supply of thiamine directs the extension of tools to gain better understanding of
the activity of the thiamine requiring enzymes of the cycle spatial aspects of metabolic regulation [79] and develop-
in a manner which controls its flux. ment of approaches to provide higher resolution infor-
mation concerning metabolic fluxes through the cycle
Inferences from the regulation of the TCA [7,80]. We firmly believe that on the application of these
cycle in non-plant systems and future tools we will finally be able to complete the jigsaw allowing
perspectives us to address the molecular hierarchy of the metabolic
As alluded to above, a further mode of regulation is that regulation of the plant mitochondrial TCA cycle.
played by compartmentation whilst studies in plants have
provided a fairly comprehensive view as to how this is Acknowledgements
controlled at the organellar level as yet little is known Financial support from the Max-Planck-Society (to WLA, ANN and ARF),
the Deutsche Forschungsgemeinschaft (grant no. DFG-SFB429 to ARF),
there in plants at finer sub-compartment resolution. This and the National Council for Scientific and Technological Development
is in sharp contrast to mammalian and microbial systems. CNPq-Brazil (grant numbers 478261/2010-1 to ANN and 472787/2011-0 to
Indeed the coining of the term metabolon to define WLA) are gratefully acknowledged.
supermolecular complexes composed of sequential meta-
bolic enzymes was first used in the context of the rat TCA References and recommended reading
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enzymes are structurally connected in metabolon result-
! of special interest
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mechanistic process for the direct delivery of a reaction
intermediate from the active site of one enzyme to the
other(s) without prior dissociation into the bulk solvent.
Metabolite channeling offers a means for high metabolic Appendix A. Supplementary data
rates with low bulk concentrations of intermediates and Supplementary data associated with this article can be
extremely rapid responses to changes in the metabolic found, in the online version, at
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