Probing in vivo Mn²⁺ speciation and oxidative stress resistance in yeast cells with electron-nuclear double resonance spectroscopy

Manganese is an essential transition metal that, among other functions, can act independently of proteins to either defend against or promote oxidative stress and disease. The majority of cellular manganese exists as low molecular-weight Mn²⁺ complexes, and the balance between opposing "essenti...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2010-08, Vol.107 (35), p.15335-15339
Main Authors: McNaughton, Rebecca L, Reddi, Amit R, Clement, Matthew H.S, Sharma, Ajay, Barnese, Kevin, Rosenfeld, Leah, Gralla, Edith Butler, Valentine, Joan Selverstone, Culotta, Valeria C, Hoffman, Brian M
Format: Article
Language:English
Subjects:
Algorithms
Biochemistry
Biological Sciences
Chromium
Electron Spin Resonance Spectroscopy - methods
Genetic engineering
Homeostasis
Kinetics
Ligands
Manganese
Manganese - chemistry
Manganese - metabolism
Metals
Models, Chemical
Mutants
Mutation
orthophosphate
Oxidative Stress
Oxygen
Oxygen - metabolism
Phosphate
Phosphates
Phosphates - chemistry
Phosphates - metabolism
Physical Sciences
Proteins
Saccharomyces cerevisiae
Saccharomyces cerevisiae - chemistry
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Speciation
Spectrophotometry, Atomic
Spectroscopy
Superoxide dismutase
Superoxide Dismutase - metabolism
Superoxides
Transition metals
Yeasts
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Summary:Manganese is an essential transition metal that, among other functions, can act independently of proteins to either defend against or promote oxidative stress and disease. The majority of cellular manganese exists as low molecular-weight Mn²⁺ complexes, and the balance between opposing "essential" and "toxic" roles is thought to be governed by the nature of the ligands coordinating Mn²⁺. Until now, it has been impossible to determine manganese speciation within intact, viable cells, but we here report that this speciation can be probed through measurements of ¹H and ³¹P electron-nuclear double resonance (ENDOR) signal intensities for intracellular Mn²⁺. Application of this approach to yeast (Saccharomyces cerevisiae) cells, and two pairs of yeast mutants genetically engineered to enhance or suppress the accumulation of manganese or phosphates, supports an in vivo role for the orthophosphate complex of Mn²⁺ in resistance to oxidative stress, thereby corroborating in vitro studies that demonstrated superoxide dismutase activity for this species.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1009648107