The molybdenum storage protein

Brünle, Steffen; Poppe, J.; Hail, Ron; Demmer, Ulrike

doi:10.1016/j.jinorgbio.2018.09.011

Cited by 8 publications

(9 citation statements)

References 28 publications

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“…decreased deuterium uptake for segment β127 to β142, indicating continuous POM cluster assembly (11).…”

Section: Resultsmentioning

confidence: 98%

“…Interestingly, the N-terminal extension also plays a crucial role in the related acetylglutamate kinase by linking dimers to higher oligomers (20). Finally, a decreased HDX was identified in segment β115 to β131 containing the glycine-rich loop β127 to β132, involved in POM cluster binding, which becomes occluded during continuous POM cluster assembly (11). Mo storage was then investigated by a reversed order of ATP and molybdate addition.…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant MoSto and MoSto Variant Production. The production of recombinant MoSto of A. vinelandii was performed in the Escherichia coli strain BL21(DE3) CC5, in which a pET21a-based vector containing the genes encoding for the MoSto αand β-subunits and a strepII-tag were incorporated (11). MoSto was purified using strep-tag affinity, anion exchange, and size exclusion chromatography and stored in the MoSto funct state in 50 mM MOPS/NaOH pH 6.5 and 50 mM NaCl at −20°C.…”

Section: Methodsmentioning

confidence: 99%

“…1). The interior of the cage serves as a container for up to approximately 12 POM clusters, whose structures are determined by specific pockets inside the cage, the 3-fold symmetry, and the preparation conditions (10,11). The polynuclear Mo-O aggregates that are covalently or noncovalently linked with the polypeptide (8,10) vary between 3 and 14 Mo atoms and are termed Mo3, Mo8, Mo5-7, hexagonal (bi)pyramidal Mo7-8, and Mo8-14 clusters ( Fig.…”

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confidence: 99%

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Molybdate pumping into the molybdenum storage protein via an ATP-powered piercing mechanism

Brünle

Eisinger

Poppe

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The molybdenum storage protein (MoSto) deposits large amounts of molybdenum as polyoxomolybdate clusters in a heterohexameric (αβ)3cage-like protein complex under ATP consumption. Here, we suggest a unique mechanism for the ATP-powered molybdate pumping process based on X-ray crystallography, cryoelectron microscopy, hydrogen-deuterium exchange mass spectrometry, and mutational studies of MoSto fromAzotobacter vinelandii. First, we show that molybdate, ATP, and Mg2+consecutively bind into the open ATP-binding groove of the β-subunit, which thereafter becomes tightly locked by fixing the previously disordered N-terminal arm of the α-subunit over the β-ATP. Next, we propose a nucleophilic attack of molybdate onto the γ-phosphate of β-ATP, analogous to the similar reaction of the structurally related UMP kinase. The formed instable phosphoric-molybdic anhydride becomes immediately hydrolyzed and, according to the current data, the released and accelerated molybdate is pressed through the cage wall, presumably by turning aside the Metβ149 side chain. A structural comparison between MoSto and UMP kinase provides valuable insight into how an enzyme is converted into a molecular machine during evolution. The postulated direct conversion of chemical energy into kinetic energy via an activating molybdate kinase and an exothermic pyrophosphatase reaction to overcome a proteinous barrier represents a novelty in ATP-fueled biochemistry, because normally, ATP hydrolysis initiates large-scale conformational changes to drive a distant process.

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“…decreased deuterium uptake for segment β127 to β142, indicating continuous POM cluster assembly (11).…”

Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Molybdate pumping into the molybdenum storage protein via an ATP-powered piercing mechanism

Brünle

Eisinger

Poppe

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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“…Although MoSto has been biochemically characterized (Brünle et al, 2018), there were no reports of MoSto activity in the in vitro FeMo-co synthesis assays. This thesis shows that Mo stored at MoSto is directly available for FeMo-co synthesis in vitro and that interaction of MoSto with FeMo-co biosynthetic proteins stimulates cofactor synthesis (Navarro-Rodríguez et al, 2019).…”

Section: Interaction Of Mosto and Nifq In Mitochondria Of S Cerevisimentioning

confidence: 99%

Molybdenum Metabolism in Azotobacter vinelandii and its Biotechnological Applications

Rodríguez¹

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Baars

Aziz

Poppe

et al. 2023

Encyclopedia of Inorganic and Bioinorganic Chemistry

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For synthesing the FeMo cofactor of nitrogenase N 2 ‐fixing bacteria continuously demand molybdate nonconstantly available in their habitats. Significant selection advantages have those species containing the molybdenum storage protein (MoSto) due to its capability to deposit ca. 130 Mo compactly as polynuclear Mo(VI)‐O or polyoxometalate (POM) clusters. Due to similar physicochemical properties, W can be stored in a related fashion. Heterododecameric MoSto is built up of two heterohexameric (αβ) 3 cages each containing 5 distinct locations for POM clusters. The complex interplay of self‐assembly and protein‐promoted processes creates a great diversity of POM clusters varying between 2 and 16 Mo atoms. Starting from smaller core species multiply linked with protein templates larger aggregates are synthesized that contain as a component an archetypical POM cluster found in solution. Mechanistically, POM cluster formation proceeds within a two‐step process. Molybdate is, at first, pressed through the cage shell in an ATP‐consuming fashion and then spontaneously polymerizes in the cage to POM clusters. Molybdate appears to be activated in the first step by phosphorylation in an ATP kinase reaction. The formed instable phosphoric‐molybdic anhydride intermediate is afterward, instantaneously, hydrolyzed in a locked chamber and the energized molybdate penetrates the protein shell at a thin place.

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The molybdenum storage protein — A bionanolab for creating experimentally alterable polyoxomolybdate clusters

Cited by 8 publications

References 28 publications

Molybdate pumping into the molybdenum storage protein via an ATP-powered piercing mechanism

Molybdate pumping into the molybdenum storage protein via an ATP-powered piercing mechanism

Molybdenum Metabolism in Azotobacter vinelandii and its Biotechnological Applications

The Molybdenum Storage Protein

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