The Mycobacterium tuberculosis Secreted Protein Rv0203 Transfers Heme to Membrane Proteins MmpL3 and MmpL11

Owens, Cedric P.; Chim, Nicholas; Graves, Amanda B.; Harmston, Christine A.; Iniguez, Angelina; Contreras, Heidi; Liptak, Matthew D.; Goulding, Celia W.

doi:10.1074/jbc.m113.453076

Cited by 73 publications

(80 citation statements)

References 73 publications

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“…That this is due to the proposed role of MmpL3 in heme iron import (40,41) is unlikely, at least in vitro, given the ability of MmpL3-DUC to produce mycobactins and, thus, to use the ferric ion present in the culture media used in this study.…”

Section: Discussionmentioning

confidence: 99%

Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

Obregón-Henao

Wallach

et al. 2016

Antimicrob Agents Chemother

103

122

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Section: Discussionmentioning

confidence: 99%

Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

Obregón-Henao

Wallach

et al. 2016

Antimicrob Agents Chemother

103

122

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“…The final purification step was performed by using an S200 10/30 Superdex column (GE Healthcare) equilibrated with a solution containing 50 mM Tris (pH 7.4) and 150 mM NaCl to yield 100% homogeneous Hal N . The percentage of heme was determined by using the pyridine hemochrome and Lowry assays (Bio-Rad) (64,65). To achieve fully heme-reconstituted Hal N , a hemin solution was added to a 1:1 molar ratio of heme to protein (Sigma) (64,65).…”

Section: Methodsmentioning

confidence: 99%

“…The percentage of heme was determined by using the pyridine hemochrome and Lowry assays (Bio-Rad) (64,65). To achieve fully heme-reconstituted Hal N , a hemin solution was added to a 1:1 molar ratio of heme to protein (Sigma) (64,65). Crystals of heme-Hal N were grown at 90 mg/ml by a hanging-drop vapor diffusion method with a reservoir containing 0.2 M ammonium sulfate, 0.1 M BisTris (pH 5.5), and 21% polyethylene glycol 3350 (PEG 3350) with a 1:1 (vol/vol) protein-to-reservoir drop.…”

Section: Methodsmentioning

confidence: 99%

Progress toward the Development of a NEAT Protein Vaccine for Anthrax Disease

et al. 2016

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Bacillus anthracis is a sporulating Gram-positive bacterium that is the causative agent of anthrax and a potential weapon of bioterrorism. The U.S.-licensed anthrax vaccine is made from an incompletely characterized culture supernatant of a nonencapsulated, toxigenic strain (anthrax vaccine absorbed [AVA]) whose primary protective component is thought to be protective antigen (PA). AVA is effective in protecting animals and elicits toxin-neutralizing antibodies in humans, but enthusiasm is dampened by its undefined composition, multishot regimen, recommended boosters, and potential for adverse reactions. Improving next-generation anthrax vaccines is important to safeguard citizens and the military. Here, we report that vaccination with recombinant forms of a conserved domain (near-iron transporter [NEAT]), common in Gram-positive pathogens, elicits protection in a murine model of B. anthracis infection. Protection was observed with both Freund's and alum adjuvants, given subcutaneously and intramuscularly, respectively, with a mixed composite of NEATs. Protection correlated with an antibody response against the NEAT domains and a decrease in the numbers of bacteria in major organs. Anti-NEAT antibodies promote opsonophagocytosis of bacilli by alveolar macrophages. To guide the development of inactive and safe NEAT antigens, we also report the crystal structure of one of the NEAT domains (Hal) and identify critical residues mediating its heme-binding and acquisition activity. These results indicate that we should consider NEAT proteins in the development of an improved antianthrax vaccine.

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“…Deletion of AnsP2 does not eliminate nitrogen assimilation from asparagine, suggesting that another protein(s) compensates for loss of function. By analogy with the recent identification of the mycobacterial corrinoid (Gopinath et al 2013b) and heme (Tullius et al 2011;Owens et al 2013) transporters, this observation reinforces the need to identify the mechanisms that mediate uptake of essential nutrients, cofactors, and trace elements in M. tuberculosis and simultaneously highlights the unique adaptation of conserved proteins to specialist functions in an obligate pathogen (discussed below). In addition, the dual role of the ansA-encoded asparaginase in mycobacterial nitrogen homeostasis and the defense against host-mediated acid stress reinforces the intersection of physiology and virulence in core metabolic functions.…”

Section: Investigating Mycobacterial Metabolism In Vivo Auxotrophs Asmentioning

confidence: 97%

Mycobacterium tuberculosis Metabolism

Warner¹

2014

Cold Spring Harbor Perspectives in Medicine

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Metabolism underpins the physiology and pathogenesis of Mycobacterium tuberculosis. However, although experimental mycobacteriology has provided key insights into the metabolic pathways that are essential for survival and pathogenesis, determining the metabolic status of bacilli during different stages of infection and in different cellular compartments remains challenging. Recent advances-in particular, the development of systems biology tools such as metabolomics-have enabled key insights into the biochemical state of M. tuberculosis in experimental models of infection. In addition, their use to elucidate mechanisms of action of new and existing antituberculosis drugs is critical for the development of improved interventions to counter tuberculosis. This review provides a broad summary of mycobacterial metabolism, highlighting the adaptation of M. tuberculosis as specialist human pathogen, and discusses recent insights into the strategies used by the host and infecting bacillus to influence the outcomes of the host -pathogen interaction through modulation of metabolic functions.Ultimately, the process of pathogenicity itself will be describable in terms of the chemistry of the mycobacterial cell.-Wheeler and Ratledge (1994) T he term "metabolism" has a very wide purview. For bacteria, it is commonly used to describe the full set of complex and interconnected chemical transformations that enable individual cells to survive and replicate. In turn, these might be broadly divided into the anabolic pathways that consume energy in generating biomass and the energy-releasing catabolic reactions that channel organic building blocks into diverse cellular structures and pathways. Metabolism also includes pathways that are essential to the ability of bacteria to respond to dynamic environments and to maintain structural integrity in the face of fluctuating nutrient availability. Therefore, in an obligate pathogen such as Mycobacterium tuberculosis whose entire life cycle is driven in the context of human infection, metabolism necessarily underpins both physiology and pathogenesis (Rhee 2013).Owing to this dual role, the metabolism of M. tuberculosis has been the subject of intense research Beste and McFadden 2010). However, although experimental mycobacteriology-in particular, the use of genetic tools to disrupt enzymatic and regulatory functions-has provided critical insights into the metabolic pathways that are esEditors:

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The Mycobacterium tuberculosis Secreted Protein Rv0203 Transfers Heme to Membrane Proteins MmpL3 and MmpL11

Cited by 73 publications

References 73 publications

Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

Progress toward the Development of a NEAT Protein Vaccine for Anthrax Disease

Mycobacterium tuberculosis Metabolism

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