Exploring the microbial metalloproteome using MIRAGE

Sevcenco, Ana–Maria; Pinkse, Martijn W. H.; Wolterbeek, Hubert Th.; Verhaert, Peter; Hagen, Wilfred R.; Hagedoorn, Peter‐Leon

doi:10.1039/c1mt00154j

Cited by 34 publications

(22 citation statements)

References 28 publications

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“…In both E. coli and Bacillus anthracis, the largest pools of protein-associated iron are in iron-superoxide dismutase (Fe-SOD) and oxidized to ferric hydroxide within ferritins and mini-ferritins (Dps family proteins) (22,23). Once oxidized, stored iron no longer contributes to the labile iron pool.…”

Section: Metal Ion Speciation In the Cellmentioning

confidence: 99%

Specificity of Metal Sensing: Iron and Manganese Homeostasis in Bacillus subtilis

Helmann

2014

Journal of Biological Chemistry

131

127

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Metalloregulatory proteins allow cells to sense metal ions and appropriately adjust the expression of metal uptake, storage, and efflux pathways. Bacillus subtilis provides a model for the coordinate regulation of iron and manganese homeostasis that involves three key regulators: Fur senses iron sufficiency, MntR senses manganese sufficiency, and PerR senses the intracellular Fe/Mn ratio. Here, I review the structural and physiological bases of selective metal perception, the effects of non-cognate metals, and mechanisms that may serve to coordinate iron and manganese homeostasis. Manganese and Iron: Chemically Similar Elements with Distinct Roles in Cell PhysiologyManganese and iron are both predominantly present as divalent cations in the reducing environment of the cytoplasm and are maintained at overall concentrations (averaged over the cell) that can approach 0.5 mM (1-3). Much of this manganese and iron is bound by enzymes that acquire metal from a kinetically accessible (labile) pool buffered in the low M range (4 -6). These two metals are not only important from the perspective of individual microbial cells, but have a global impact due to their central roles in photosynthesis, nitrogen fixation, and other key steps in elemental cycling.Nearly all cellular life requires significant amounts of iron. The sole documented exceptions are the lactobacilli and the spirochete Borrelia burgdorferi, which have little if any requirement for iron (3). Coincident with this disregard for iron, these same organisms have high requirements for manganese and have been described as manganese-centric (7). Conversely, other organisms have little demonstrable requirement for manganese, although they may use manganese when available. The best characterized is Escherichia coli, which has a largely ironcentric metabolism, but conditionally imports manganese in response to oxidative stress (8).Here, I review the key factors regulating iron and Mn homeostasis in the model organism Bacillus subtilis. B. subtilis encodes three members of the Fur 2 family (Fur, PerR, and Zur) and one member of the DtxR/MntR family (MntR) that regulate the import of nutrient metal ions (9 -11). Structural and biochemical studies have provided insights into the mechanisms of selective activation of these repressors by their cognate metal ions (12-14). B. subtilis requires both iron and manganese for growth, a property shared with many pathogens for which the ability to obtain these metals from the host is a critical determinant of virulence (7,(15)(16)(17). Metalloregulators as a Window into Cellular PhysiologyMetalloregulators function as the arbiters of metal ion sufficiency and excess (6,12,18). For those that sense sufficiency, the affinity (measured as K d ) for their cognate metal(s) defines the level of metal judged to be sufficient. When free metal concentrations rise above this level, the regulator transitions from its inactive (apo-) form to its activated (holo-) form and represses expression of metal import. In B. subtilis, Fur serves as ...

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Section: Metal Ion Speciation In the Cellmentioning

confidence: 99%

Specificity of Metal Sensing: Iron and Manganese Homeostasis in Bacillus subtilis

Helmann

2014

Journal of Biological Chemistry

131

127

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“…One approach that exploits the use of metal radionuclides is Metal Isotope native RadioAutography in Gel Electrophoresis (MIRAGE) [12,18]. A micro-organism is grown in a medium spiked with a particular metal radionuclide.…”

Section: Metalloproteomic Approachesmentioning

confidence: 99%

“…The metalloproteomes of several bacteria and archaea have been studied in significant detail [10,11,12,13,14,15,16]. Major metalloproteomic methods are presented, important research findings that have been obtained from metalloproteomics research are discussed, and finally, an outlook is given on emerging approaches that may propel this field in the near future.…”

Section: Introductionmentioning

confidence: 99%

Microbial Metalloproteomics

Hagedoorn

2015

Proteomes

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Metalloproteomics is a rapidly developing field of science that involves the comprehensive analysis of all metal-containing or metal-binding proteins in a biological sample. The purpose of this review is to offer a comprehensive overview of the research involving approaches that can be categorized as inductively coupled plasma (ICP)-MS based methods, X-ray absorption/fluorescence, radionuclide based methods and bioinformatics. Important discoveries in microbial proteomics will be reviewed, as well as the outlook to new emerging approaches and research areas.

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“…This work was performed by first labeling the proteins with a short-lived isotope, separating them, detecting them by autoradiography, and then identifying the proteins by mass spectrometry. This approach has been used to work to identify the major copper-binding proteins in the periplasm of E. coli , and in work to identify the soluble iron and zinc proteins in the same organism [46,47]. An advantage of this technique, as with liquid chromatography/ICP-MS approaches, is that pulse-chase experiments to identify newly-formed protein are possible.…”

Section: Survey Of Approaches To Metalloproteomicsmentioning

confidence: 99%

Metalloproteomics: challenges and prospective for clinical research applications

Fu¹,

Finney

2014

Expert Review of Proteomics

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Metals are essential cofactors, utilized in many critical cellular processes. For example, zinc is important in insulin biosynthesis, and may play a role in Alzheimer's disease, but much of how this happens remains unknown. Knowing which metal is in which protein at a given point in time would lead to new insights into how they do their work. New tools are being developed to investigate this, with potential for biomedical applications. In this report, we consider the promise and limitations of these techniques. We provide a brief overview of the techniques available, and a discussion of the technical challenges to biomedical applications, with particular focus on what must be overcome for the potential of these approaches to be achieved.

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Exploring the microbial metalloproteome using MIRAGE

Cited by 34 publications

References 28 publications

Specificity of Metal Sensing: Iron and Manganese Homeostasis in Bacillus subtilis

Specificity of Metal Sensing: Iron and Manganese Homeostasis in Bacillus subtilis

Microbial Metalloproteomics

Metalloproteomics: challenges and prospective for clinical research applications

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