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von Ballmoos, Christoph
Publikationer (10 of 22) Visa alla publikationer
Lundgren, C. A. K., Sjöstrand, D., Biner, O., Bennett, M., Rudling, A., Johansson, A.-L., . . . Högbom, M. (2018). Scavenging of superoxide by a membrane-bound superoxide oxidase. Nature Chemical Biology, 14, 788-793
Öppna denna publikation i ny flik eller fönster >>Scavenging of superoxide by a membrane-bound superoxide oxidase
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2018 (Engelska)Ingår i: Nature Chemical Biology, ISSN 1552-4450, E-ISSN 1552-4469, Vol. 14, s. 788-793Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Superoxide is a reactive oxygen species produced during aerobic metabolism in mitochondria and prokaryotes. It causes damage to lipids, proteins and DNA and is implicated in cancer, cardiovascular disease, neurodegenerative disorders and aging. As protection, cells express soluble superoxide dismutases, disproportionating superoxide to oxygen and hydrogen peroxide. Here, we describe a membrane-bound enzyme that directly oxidizes superoxide and funnels the sequestered electrons to ubiquinone in a diffusion-limited reaction. Experiments in proteoliposomes and inverted membranes show that the protein is capable of efficiently quenching superoxide generated at the membrane in vitro. The 2.0 Å crystal structure shows an integral membrane di-heme cytochrome b poised for electron transfer from the P-side and proton uptake from the N-side. This suggests that the reaction is electrogenic and contributes to the membrane potential while also conserving energy by reducing the quinone pool. Based on this enzymatic activity, we propose that the enzyme family be denoted superoxide oxidase (SOO).

Nationell ämneskategori
Biologiska vetenskaper
Forskningsämne
biokemi med inriktning mot bioinformatik; biokemi
Identifikatorer
urn:nbn:se:su:diva-156235 (URN)10.1038/s41589-018-0072-x (DOI)000438970200013 ()
Tillgänglig från: 2018-05-03 Skapad: 2018-05-03 Senast uppdaterad: 2022-02-26Bibliografiskt granskad
von Ballmoos, C., Smirnova, I., Poiana, F., Gonska, N., Chang, H.-Y., Gennis, R. B., . . . Ädelroth, P. (2017). Dynamics of the K-B Proton Pathway in Cytochrome ba(3) from Thermus thermophilus. Israel Journal of Chemistry, 57(5), 424-436
Öppna denna publikation i ny flik eller fönster >>Dynamics of the K-B Proton Pathway in Cytochrome ba(3) from Thermus thermophilus
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2017 (Engelska)Ingår i: Israel Journal of Chemistry, ISSN 0021-2148, Vol. 57, nr 5, s. 424-436Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The ba(3) cytochrome c oxidase from Thermus thermophilus is a B-type oxygen-reducing heme-copper oxidase and a proton pump. It uses only one proton pathway for transfer of protons to the catalytic site, the K-B pathway. It was previously shown that the ba(3) oxidase has an overall similar reaction sequence to that in mitochondrial-like A-type oxidases. However, the timing of loading the pump site, and formation and decay of catalytic intermediates is different in the two types of oxidases. In the present study, we have investigated variants in which two amino acids of the K-B proton pathway leading to the catalytic site were exchanged; Tyr-248 (located approximate to 23 angstrom below the active site towards the cytoplasm) in subunit I (Y248T) and Glu-15 (approximate to 26 angstrom below the active site, approximate to 16 angstrom from Tyr-248) in subunit II (E15(II)Q). Even though the overall catalytic turnover in these two variants is similar and very low (<1% of wildtype), the substitutions had distinctly different effects on the kinetics of proton transfer to the catalytic site. The results indicate that the Glu-15(II) is the only essentially crucial residue of the K-B pathway, but that the Tyr-248 also plays a distinct role in defining an internal proton donor and controlling the dynamics of proton transfer to the pump site and the catalytic site.

Nyckelord
heme-copper oxidases, cytochrome c oxidase, proton transfer, electron transfer, membrane protein, respiration, redox reaction, metalloprotein, cytochrome aa(3), cytochrome cbb(3)
Nationell ämneskategori
Biologiska vetenskaper
Forskningsämne
biokemi
Identifikatorer
urn:nbn:se:su:diva-144708 (URN)10.1002/ijch.201600136 (DOI)000401329000009 ()
Tillgänglig från: 2017-07-21 Skapad: 2017-07-21 Senast uppdaterad: 2022-02-28Bibliografiskt granskad
Nilsson, T., Rydström Lundin, C., Nordlund, G., Ädelroth, P., von Ballmoos, C. & Brzezinski, P. (2016). Lipid-mediated Protein-protein Interactions Modulate Respiration-driven ATP Synthesis. Scientific Reports, 6, Article ID 24113.
Öppna denna publikation i ny flik eller fönster >>Lipid-mediated Protein-protein Interactions Modulate Respiration-driven ATP Synthesis
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2016 (Engelska)Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 6, artikel-id 24113Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Energy conversion in biological systems is underpinned by membrane-bound proton transporters that generate and maintain a proton electrochemical gradient across the membrane which used, e.g. for generation of ATP by the ATP synthase. Here, we have co-reconstituted the proton pump cytochrome bo3 (ubiquinol oxidase) together with ATP synthase in liposomes and studied the effect of changing the lipid composition on the ATP synthesis activity driven by proton pumping. We found that for 100 nm liposomes, containing 5 of each proteins, the ATP synthesis rates decreased significantly with increasing fractions of DOPA, DOPE, DOPG or cardiolipin added to liposomes made of DOPC; with e.g. 5% DOPG, we observed an almost 50% decrease in the ATP synthesis rate. However, upon increasing the average distance between the proton pumps and ATP synthases, the ATP synthesis rate dropped and the lipid dependence of this activity vanished. The data indicate that protons are transferred along the membrane, between cytochrome bo3 and the ATP synthase, but only at sufficiently high protein densities. We also argue that the local protein density may be modulated by lipid-dependent changes in interactions between the two proteins complexes, which points to a mechanism by which the cell may regulate the overall activity of the respiratory chain.

Nationell ämneskategori
Biologiska vetenskaper
Forskningsämne
biokemi
Identifikatorer
urn:nbn:se:su:diva-130172 (URN)10.1038/srep24113 (DOI)000374219300001 ()27063297 (PubMedID)
Tillgänglig från: 2016-05-11 Skapad: 2016-05-09 Senast uppdaterad: 2022-09-15Bibliografiskt granskad
Lindholm, L., Ariöz, C., Jawurek, M., Liebau, J., Mäler, L., Wieslander, Å., . . . Barth, A. (2015). Effect of lipid bilayer properties on the photocycle of green proteorhodopsin. Biochimica et Biophysica Acta - Bioenergetics, 1847(8), 698-708
Öppna denna publikation i ny flik eller fönster >>Effect of lipid bilayer properties on the photocycle of green proteorhodopsin
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2015 (Engelska)Ingår i: Biochimica et Biophysica Acta - Bioenergetics, ISSN 0005-2728, E-ISSN 1879-2650, Vol. 1847, nr 8, s. 698-708Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The significance of specific lipids for proton pumping by the bacterial rhodopsin proteorhodopsin (pR) was studied. To this end, it was examined whether pR preferentially binds certain lipids and whether molecular properties of the lipid environment affect the photocycle. pR's photocyde was followed by microsecond flash-photolysis in the visible spectral range. It was fastest in phosphatidylcholine liposomes (soy bean lipid), intermediate in 3-[(3-cholamidopropyl) dimethylammonio] propanesulfonate (CHAPS): 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bicelles and in Triton X-100, and slowest when pR was solubilized in CHAPS. In bicelles with different lipid compositions, the nature of the head groups, the unsaturation level and the fatty acid chain length had small effects on the photocycle. The specific affinity of pR for lipids of the expression host Eschetichia coil was investigated by an optimized method of lipid isolation from purified membrane protein using two different concentrations of the detergent N-dodecyl-beta-D-maltoside (DDM). We found that 11 lipids were copurified per pR molecule at 0.1% DDM, whereas essentially all lipids were stripped off from pR by 1% DDM. The relative amounts of copurifled phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin did not correlate with the molar percentages normally present in E. coil cells. The results indicate a predominance of phosphatidylethanolamine species in the lipid annulus around recombinant pR that are less polar than the dominant species in the cell membrane of the expression host E. coli.

Nyckelord
Proteorhodopsin, Bicelle, Lipid, Detergent, Membrane protein, Photocycle
Nationell ämneskategori
Biologiska vetenskaper
Identifikatorer
urn:nbn:se:su:diva-119116 (URN)10.1016/j.bbabio.2015.04.011 (DOI)000356547100003 ()
Tillgänglig från: 2015-07-31 Skapad: 2015-07-29 Senast uppdaterad: 2022-02-23Bibliografiskt granskad
von Ballmoos, C., Gonska, N., Lachmann, P., Gennis, R. B., Ädelroth, P. & Brzezinski, P. (2015). Mutation of a single residue in the ba(3) oxidase specifically impairs protonation of the pump site. Proceedings of the National Academy of Sciences of the United States of America, 112(11), 3397-3402
Öppna denna publikation i ny flik eller fönster >>Mutation of a single residue in the ba(3) oxidase specifically impairs protonation of the pump site
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2015 (Engelska)Ingår i: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 112, nr 11, s. 3397-3402Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The ba(3)-type cytochrome c oxidase from Thermus thermophilus is a membrane-bound protein complex that couples electron transfer to O-2 to proton translocation across the membrane. To elucidate the mechanism of the redox-driven proton pumping, we investigated the kinetics of electron and proton transfer in a structural variant of the ba(3) oxidase where a putative pump site was modified by replacement of Asp372 by Ile. In this structural variant, proton pumping was uncoupled from internal electron transfer and O-2 reduction. The results from our studies show that proton uptake to the pump site (time constant similar to 65 mu s in the wild-type cytochrome c oxidase) was impaired in the Asp372Ile variant. Furthermore, a reaction step that in the wild-type cytochrome c oxidase is linked to simultaneous proton uptake and release with a time constant of similar to 1.2 ms was slowed to similar to 8.4 ms, and in Asp372Ile was only associated with proton uptake to the catalytic site. These data identify reaction steps that are associated with protonation and deprotonation of the pump site, and point to the area around Asp372 as the location of this site in the ba(3) cytochrome c oxidase.

Nyckelord
cytochrome c oxidase, membrane protein, respiration, cytochrome aa(3), electron transfer
Nationell ämneskategori
Biologiska vetenskaper
Forskningsämne
biokemi
Identifikatorer
urn:nbn:se:su:diva-116611 (URN)10.1073/pnas.1422434112 (DOI)000351060000072 ()25733886 (PubMedID)
Anmärkning

AuthorCount:6;

Tillgänglig från: 2015-04-30 Skapad: 2015-04-22 Senast uppdaterad: 2022-02-23Bibliografiskt granskad
Lee, C., Yashiro, S., Dotson, D. L., Uzdavinys, P., Iwata, S., Sansom, M. S. P., . . . Cameron, A. D. (2014). Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights. The Journal of General Physiology, 144(6), 529-544
Öppna denna publikation i ny flik eller fönster >>Crystal structure of the sodium-proton antiporter NhaA dimer and new mechanistic insights
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2014 (Engelska)Ingår i: The Journal of General Physiology, ISSN 0022-1295, E-ISSN 1540-7748, Vol. 144, nr 6, s. 529-544Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Sodium-proton antiporters rapidly exchange protons and sodium ions across the membrane to regulate intracellular pH, cell volume, and sodium concentration. How ion binding and release is coupled to the conformational changes associated with transport is not clear. Here, we report a crystal form of the prototypical sodium-proton antiporter NhaA from Escherichia coli in which the protein is seen as a dimer. In this new structure, we observe a salt bridge between an essential aspartic acid (Asp163) and a conserved lysine (Lys300). An equivalent salt bridge is present in the homologous transporter NapA, but not in the only other known crystal structure of NhaA, which provides the foundation of most existing structural models of electrogenic sodium-proton antiport. Molecular dynamics simulations show that the stability of the salt bridge is weakened by sodium ions binding to Asp164 and the neighboring Asp163. This suggests that the transport mechanism involves Asp163 switching between forming a salt bridge with Lys300 and interacting with the sodium ion. pK(a) calculations suggest that Asp163 is highly unlikely to be protonated when involved in the salt bridge. As it has been previously suggested that Asp163 is one of the two residues through which proton transport occurs, these results have clear implications to the current mechanistic models of sodium-proton antiport in NhaA.

Nationell ämneskategori
Biologiska vetenskaper
Forskningsämne
biokemi
Identifikatorer
urn:nbn:se:su:diva-111902 (URN)10.1085/jgp.201411219 (DOI)000345565900006 ()
Anmärkning

AuthorCount:10;

Tillgänglig från: 2015-01-13 Skapad: 2015-01-08 Senast uppdaterad: 2022-03-23Bibliografiskt granskad
Nordlund, G., Brzezinski, P. & von Ballmoos, C. (2014). SNARE-fusion mediated insertion of membrane proteins into native and artificial membranes. Nature Communications, 5, 4303
Öppna denna publikation i ny flik eller fönster >>SNARE-fusion mediated insertion of membrane proteins into native and artificial membranes
2014 (Engelska)Ingår i: Nature Communications, E-ISSN 2041-1723, Vol. 5, s. 4303-Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Membrane proteins carry out functions such as nutrient uptake, ATP synthesis or transmembrane signal transduction. An increasing number of reports indicate that cellular processes are underpinned by regulated interactions between these proteins. Consequently, functional studies of these networks at a molecular level require co-reconstitution of the interacting components. Here, we report a SNARE protein-based method for incorporation of multiple membrane proteins into artificial membrane vesicles of well-defined composition, and for delivery of large water-soluble substrates into these vesicles. The approach is used for in vitro reconstruction of a fully functional bacterial respiratory chain from purified components. Furthermore, the method is used for functional incorporation of the entire F1F0 ATP synthase complex into native bacterial membranes from which this component had been genetically removed. The novel methodology offers a tool to investigate complex interaction networks between membrane-bound proteins at a molecular level, which is expected to generate functional insights into key cellular functions.

Nationell ämneskategori
Biologiska vetenskaper
Identifikatorer
urn:nbn:se:su:diva-107442 (URN)10.1038/ncomms5303 (DOI)000340613800011 ()
Anmärkning

AuthorCount:3;

Tillgänglig från: 2014-09-17 Skapad: 2014-09-15 Senast uppdaterad: 2023-03-28Bibliografiskt granskad
Lee, C., Kang, H. J., von Ballmoos, C., Newstead, S., Uzdavinys, P., Dotson, D. L., . . . Drew, D. (2013). A two-domain elevator mechanism for sodium/proton antiport. Nature, 501(7468), 573-577
Öppna denna publikation i ny flik eller fönster >>A two-domain elevator mechanism for sodium/proton antiport
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2013 (Engelska)Ingår i: Nature, ISSN 0028-0836, E-ISSN 1476-4687, Vol. 501, nr 7468, s. 573-577Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Sodium/proton (Na+/H+) antiporters, located at the plasma membrane in every cell, are vital for cell homeostasis1. In humans, their dysfunction has been linked to diseases, such as hypertension, heart failure and epilepsy, and they are well-established drug targets(2). The best understood model system for Na+/H+ antiport is NhaA from Escherichia coli(1,3), for which both electron microscopy and crystal structures are available(4-6). NhaA is made up of two distinct domains: a core domain and a dimerization domain. In the NhaA crystal structure a cavity is located between the two domains, providing access to the ion-binding site from the inward-facing surface of the protein(1,4). Likemany Na+/H+ antiporters, the activity of NhaA is regulated by pH, only becoming active above pH 6.5, at which point a conformational change is thought to occur(7). The only reported NhaA crystal structure so far is of the low pH inactivated form(4). Here we describe the active-state structure of a Na+/H+ antiporter, NapA from Thermus thermophilus, at 3 angstrom resolution, solved from crystals grown at pH7.8. In the NapA structure, the core and dimerization domains are in different positions to those seen in NhaA, and a negatively charged cavity has now opened to the outside. The extracellular cavity allows access to a strictly conserved aspartate residue thought to coordinate ion binding(1,8,9) directly, a role supported hereby molecular dynamics simulations. To alternate access to this ion-binding site, however, requires a surprisingly large rotation of the core domain, some 20 degrees against the dimerization interface. We conclude that despite their fast transport rates of up to 1,500 ions per second(3), Na+/H+ antiporters operate by a two-domain rocking bundle model, revealing themes relevant to secondary-active transporters in general.

Nationell ämneskategori
Biologiska vetenskaper
Forskningsämne
biokemi
Identifikatorer
urn:nbn:se:su:diva-95768 (URN)10.1038/nature12484 (DOI)000324826300064 ()
Anmärkning

AuthorCount:10;

Tillgänglig från: 2013-11-04 Skapad: 2013-11-04 Senast uppdaterad: 2022-02-24Bibliografiskt granskad
Gigliobianco, T., Gangolf, M., Lakaye, B., Pirson, B., von Ballmoos, C., Wins, P. & Bettendorff, L. (2013). An alternative role of FoF1-ATP synthase in Escherichia coli: synthesis of thiamine triphosphate. Scientific Reports, 3, 1071
Öppna denna publikation i ny flik eller fönster >>An alternative role of FoF1-ATP synthase in Escherichia coli: synthesis of thiamine triphosphate
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2013 (Engelska)Ingår i: Scientific Reports, E-ISSN 2045-2322, Vol. 3, s. 1071-Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

In E. coli, thiamine triphosphate (ThTP), a putative signaling molecule, transiently accumulates in response to amino acid starvation. This accumulation requires the presence of an energy substrate yielding pyruvate. Here we show that in intact bacteria ThTP is synthesized from free thiamine diphosphate (ThDP) and P-i, the reaction being energized by the proton-motive force (Delta p) generated by the respiratory chain. ThTP production is suppressed in strains carrying mutations in F-1 or a deletion of the atp operon. Transformation with a plasmid encoding the whole atp operon fully restored ThTP production, highlighting the requirement for FoF1-ATP synthase in ThTP synthesis. Our results show that, under specific conditions of nutritional downshift, FoF1-ATP synthase catalyzes the synthesis of ThTP, rather than ATP, through a highly regulated process requiring pyruvate oxidation. Moreover, this chemiosmotic mechanism for ThTP production is conserved from E. coli to mammalian brain mitochondria.

Nationell ämneskategori
Biologiska vetenskaper
Identifikatorer
urn:nbn:se:su:diva-88361 (URN)10.1038/srep01071 (DOI)000313551000005 ()
Anmärkning

AuthorCount:7;

Tillgänglig från: 2013-03-25 Skapad: 2013-03-13 Senast uppdaterad: 2022-09-15Bibliografiskt granskad
Oliynyk, V., Mille, C., Ng, J. B. S., von Ballmoos, C., Corkery, R. W. & Bergström, L. (2013). Selective and atp driven transport of ions across supported membranes into nanoporous carriers using gramicidin a and atp synthase. Physical Chemistry, Chemical Physics - PCCP, 15(8), 2733-2740
Öppna denna publikation i ny flik eller fönster >>Selective and atp driven transport of ions across supported membranes into nanoporous carriers using gramicidin a and atp synthase
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2013 (Engelska)Ingår i: Physical Chemistry, Chemical Physics - PCCP, ISSN 1463-9076, E-ISSN 1463-9084, Vol. 15, nr 8, s. 2733-2740Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

We report a robust and versatile membrane protein based system for selective uptake and release of ions from nanoporous particles sealed with ion-tight lipid bilayers of various compositions that is driven by the addition of ATP or a chemical potential gradient. We have successfully incorporated both a passive ion channel-type peptide (gramicidin A) and a more complex primary sodium ion transporter (ATP synthase) into the supported lipid bilayers on solid nanoporous silica particles. Protein-mediated controlled release/uptake of sodium ions across the ion-tight lipid bilayer seal from or into the nanoporous silica carrier was imaged in real time using a confocal laser scanning microscope and the intensity changes were quantified. ATP-driven transport of sodium ions across the supported lipid bilayer against a chemical gradient was demonstrated. The possibility of designing durable carriers with tight lipid membranes, containing membrane proteins for selective ion uptake and release, offers new possibilities for functional studies of single or cascading membrane protein systems and could also be used as biomimetic microreactors for controlled synthesis of inorganic multicomponent materials.

Nationell ämneskategori
Kemi Fysikalisk kemi Biokemi och molekylärbiologi
Identifikatorer
urn:nbn:se:su:diva-88325 (URN)10.1039/c2cp43166a (DOI)000314365800014 ()
Forskningsfinansiär
VetenskapsrådetVinnovaKK-stiftelsenStiftelsen för strategisk forskning (SSF)
Anmärkning

AuthorCount:6;

Tillgänglig från: 2013-03-19 Skapad: 2013-03-12 Senast uppdaterad: 2022-02-24Bibliografiskt granskad
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