Mycobacterium tuberculosis Phosphoenolpyruvate Carboxykinase Is Regulated by Redox Mechanisms and Interaction with Thioredoxin

Machová, Iva; Snášel, Jan; Zimmermann, Michael; Laubitz, Daniel; Płociński, Przemysław; Oehlmann, Wulf; Singh, Mahavir; Dostál, Jiřı́; Sauer, Uwe; Pichová, Iva

doi:10.1074/jbc.m113.536748

Cited by 29 publications

(48 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mtb Pck is a GTP-dependent enzyme that under standard conditions catalyzes the gluconeogenic reaction leading to production of PEP. However, under reducing conditions, it favors the anaplerotic OAA synthesis [ 16 ]. The role of Pck in fixation of CO 2 during OAA biosynthesis has also been reported for slowly or intracellularly growing Mtb [ 17 , 18 ] and Mtb cultivated in conditions mimicking hypoxia arrest [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Structural and Functional Studies of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis

et al. 2015

Self Cite

View full text Add to dashboard Cite

Tuberculosis, the second leading infectious disease killer after HIV, remains a top public health priority. The causative agent of tuberculosis, Mycobacterium tuberculosis (Mtb), which can cause both acute and clinically latent infections, reprograms metabolism in response to the host niche. Phosphoenolpyruvate carboxykinase (Pck) is the enzyme at the center of the phosphoenolpyruvate-pyruvate-oxaloacetate node, which is involved in regulating the carbon flow distribution to catabolism, anabolism, or respiration in different states of Mtb infection. Under standard growth conditions, Mtb Pck is associated with gluconeogenesis and catalyzes the metal-dependent formation of phosphoenolpyruvate. In non-replicating Mtb, Pck can catalyze anaplerotic biosynthesis of oxaloacetate. Here, we present insights into the regulation of Mtb Pck activity by divalent cations. Through analysis of the X-ray structure of Pck-GDP and Pck-GDP-Mn2+ complexes, mutational analysis of the GDP binding site, and quantum mechanical (QM)-based analysis, we explored the structural determinants of efficient Mtb Pck catalysis. We demonstrate that Mtb Pck requires presence of Mn2+ and Mg2+ cations for efficient catalysis of gluconeogenic and anaplerotic reactions. The anaplerotic reaction, which preferably functions in reducing conditions that are characteristic for slowed or stopped Mtb replication, is also effectively activated by Fe2+ in the presence of Mn2+ or Mg2+ cations. In contrast, simultaneous presence of Fe2+ and Mn2+ or Mg2+ inhibits the gluconeogenic reaction. These results suggest that inorganic ions can contribute to regulation of central carbon metabolism by influencing the activity of Pck. Furthermore, the X-ray structure determination, biochemical characterization, and QM analysis of Pck mutants confirmed the important role of the Phe triad for proper binding of the GDP-Mn2+ complex in the nucleotide binding site and efficient catalysis of the anaplerotic reaction.

show abstract

Section: Introductionmentioning

confidence: 99%

Structural and Functional Studies of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Dissecting M Tuberculosis Pathogenesis From Protein Interacmentioning

confidence: 98%

“…The gene expression of Trx is also increased under hypoxic conditions. These findings suggest the presence of a regulatory mechanism by which M. tuberculosis responds to hypoxia and then adjusts metabolic activity to adapt to low-oxygen environments [12].As an intracellular pathogen, M. tuberculosis encounters oxidative stress throughout its life inside the host environment. Trx is the major protein disulfide reductase responsible for maintaining the redox state of the cytosol.…”

mentioning

confidence: 97%

See 1 more Smart Citation

Improved understanding of pathogenesis from protein interactions inMycobacteriumtuberculosis

Cui

He²

2014

Expert Review of Proteomics

View full text Add to dashboard Cite

Comprehensive mapping and analysis of protein-protein interactions provide not only systematic approaches for dissecting the infection and survival mechanisms of pathogens but also clues for discovering new antibacterial drug targets. Protein interaction data on Mycobacterium tuberculosis have rapidly accumulated over the past several years. This review summarizes the current progress of protein interaction studies on M. tuberculosis, the causative agent of tuberculosis. These efforts improve our knowledge on the stress response, signaling regulation, protein secretion and drug resistance of the bacteria. M. tuberculosis-host protein interaction studies, although still limited, have recently opened a new door for investigating the pathogenesis of the bacteria. Finally, this review discusses the importance of protein interaction data on identifying and screening new anti-tuberculosis targets and drugs, respectively.KEYWORDS: drug targets • Mycobacterium tuberculosis • pathogen-host interaction • protein-protein interaction • stress response Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is one of the most devastating human pathogens. In 2011, there were an estimated 8.7 million new cases of TB and 1.4 million people died from TB [1]. Sophisticated regulations allow M. tuberculosis to counter host-exerted harmful environments, interfere with host signaling pathways, evade immune clearance, survive and replicate in the host. Intracellular protein-protein interactions serve important functions in many complex cellular activities, such as stress response, cell signaling, transcriptional regulation, cell division, cell wall synthesis and protein secretion. In addition, interspecies pathogen-host protein interactions help the pathogen to interfere with host pathways and then evade host immune killing. Analyzing the human protein interactions involved in M. tuberculosis infection provides insights into the infection process. All these kinds of protein interaction data provide a basis for us to understand the molecular mechanisms of M. tuberculosis pathogenesis as well as clues for anti-TB drug design (FIGURE 1). In 1998, Cole et al. sequenced the complete genome of M. tuberculosis and found that this pathogen encodes approximately 4000 genes [2]. Since then, many studies have analyzed the interactions of these genes by using computational and highthroughput experimental technologies.Protein interaction data resource of M. tuberculosis Protein-protein interactions include physical interactions and functional interactions, in which physical interactions refer to the direct binding of two proteins and functional interactions mean these proteins are of functional relevance, for example, they are components of a same cell pathway. Unless specified otherwise, protein interactions in this review mean physical interactions. Obtaining protein-protein interaction data is important to elucidate the functions and mechanisms of this interaction in different cellular processes. The recent advances in the computa...

show abstract

“…Thus, like the case for ICL, these studies identified pckA as a clear determinant of persistence but failed to reveal its specific metabolic role, owing to limitations in the accuracy and completeness of bio-informatic annotations and in vitro biochemical studies. Indeed, follow on biochemical studies demonstrated that M. tuberculosis ’s PckA could catalyze the reverse reaction in an anaplerotic, rather than gluconeogenic, direction under reducing conditions such as those associated with hypoxia (103). Experimentally-based computational models of metabolic flux have similarly suggested that M. tuberculosis PckA may operate in an anaplerotic direction during intracellular growth within a THP-1 macrophage-like cell line (55).…”

Section: Measurementsmentioning

confidence: 99%

Metabolic Perspectives on Persistence

Hartman¹,

Wang²,

Jansen³

et al. 2017

Microbiol Spectr

View full text Add to dashboard Cite

SUMMARY Accumulating evidence has left little doubt about the importance of persistence or metabolism in the biology and chemotherapy of tuberculosis. However, knowledge of the intersection between these two factors has begun to emerge only recently. Here, we provide a focused review of metabolic characteristics associated with M. tuberculosis persistence. We focus on metabolism because it is the biochemical foundation of all physiologic processes and a distinguishing hallmark of M. tuberculosis’s physiology and pathogenicity. In addition, it serves as the chemical interface between host and pathogen. However, existing knowledge derives largely from physiologic contexts in which replication is the primary biochemical objective. The goal of this review is to reframe existing knowledge of M. tuberculosis metabolism in the context of persistence where quiescence is often a key distinguishing characteristic. Such a perspective may help guide ongoing efforts to develop more efficient cures and inform on novel strategies to break the cycle of transmission sustaining the pandemic.

show abstract

Mycobacterium tuberculosis Phosphoenolpyruvate Carboxykinase Is Regulated by Redox Mechanisms and Interaction with Thioredoxin

Cited by 29 publications

References 34 publications

Structural and Functional Studies of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis

Structural and Functional Studies of Phosphoenolpyruvate Carboxykinase from Mycobacterium tuberculosis

Improved understanding of pathogenesis from protein interactions inMycobacteriumtuberculosis

Metabolic Perspectives on Persistence

Contact Info

Product

Resources

About