umu.sePublications
Change search
Link to record
Permanent link

Direct link
BETA
Weise, Christoph
Alternative names
Publications (10 of 12) Show all publications
Perdersen, M. N., Fodera, V., Horvath, I., van Maarschalkerweerd, A., Toft, K. N., Weise, C., . . . Vestergaard, B. (2015). Direct Correlation Between Ligand-Induced alpha-Synuclein Oligomers and Amyloid-like Fibril Growth. Scientific Reports, 5, Article ID 10422.
Open this publication in new window or tab >>Direct Correlation Between Ligand-Induced alpha-Synuclein Oligomers and Amyloid-like Fibril Growth
Show others...
2015 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 5, article id 10422Article in journal (Refereed) Published
Abstract [en]

Aggregation of proteins into amyloid deposits is the hallmark of several neurodegenerative diseases such as Alzheimer's and Parkinson's disease. The suggestion that intermediate oligomeric species may be cytotoxic has led to intensified investigations of pre-fibrillar oligomers, which are complicated by their transient nature and low population. Here we investigate alpha-synuclein oligomers, enriched by a 2-pyridone molecule (FN075), and the conversion of oligomers into fibrils. As probed by leakage assays, the FN075 induced oligomers potently disrupt vesicles in vitro, suggesting a potential link to disease related degenerative activity. Fibrils formed in the presence and absence of FN075 are indistinguishable on microscopic and macroscopic levels. Using small angle X-ray scattering, we reveal that FN075 induced oligomers are similar, but not identical, to oligomers previously observed during alpha-synuclein fibrillation. Since the levels of FN075 induced oligomers correlate with the amounts of fibrils among different FN075: protein ratios, the oligomers appear to be on-pathway and modeling supports an 'oligomer stacking model' for alpha-synuclein fibril elongation.

Place, publisher, year, edition, pages
Springer, 2015
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-106505 (URN)10.1038/srep10422 (DOI)000355546300001 ()
Available from: 2015-07-16 Created: 2015-07-14 Last updated: 2019-03-14Bibliographically approved
Weise, C., Login, F. H., Ho, O., Gröbner, G., Wolf-Watz, H. & Wolf-Watz, M. (2014). Negatively charged lipid membranes promote a disorder-order transition in the Yersinia YscU protein. Biophysical Journal, 107(8), 1950-1961
Open this publication in new window or tab >>Negatively charged lipid membranes promote a disorder-order transition in the Yersinia YscU protein
Show others...
2014 (English)In: Biophysical Journal, ISSN 0006-3495, E-ISSN 1542-0086, Vol. 107, no 8, p. 1950-1961Article in journal (Refereed) Published
Abstract [en]

The inner membrane of Gram-negative bacteria is negatively charged, rendering positively charged cytoplasmic proteins in close proximity likely candidates for protein-membrane interactions. YscU is a Yersinia pseudotuberculosis type III secretion system protein crucial for bacterial pathogenesis. The protein contains a highly conserved positively charged linker sequence that separates membrane-spanning and cytoplasmic (YscUC) domains. Although disordered in solution, inspection of the primary sequence of the linker reveals that positively charged residues are separated with a typical helical periodicity. Here, we demonstrate that the linker sequence of YscU undergoes a largely electrostatically driven coil-to-helix transition upon binding to negatively charged membrane interfaces. Using membrane-mimicking sodium dodecyl sulfate micelles, an NMR derived structural model reveals the induction of three helical segments in the linker. The overall linker placement in sodium dodecyl sulfate micelles was identified by NMR experiments including paramagnetic relaxation enhancements. Partitioning of individual residues agrees with their hydrophobicity and supports an interfacial positioning of the helices. Replacement of positively charged linker residues with alanine resulted in YscUC variants displaying attenuated membrane-binding affinities, suggesting that the membrane interaction depends on positive charges within the linker. In vivo experiments with bacteria expressing these YscU replacements resulted in phenotypes displaying significantly reduced effector protein secretion levels. Taken together, our data identify a previously unknown membrane-interacting surface of YscUC that, when perturbed by mutations, disrupts the function of the pathogenic machinery in Yersinia.

Place, publisher, year, edition, pages
Cell Press, 2014
National Category
Biophysics
Identifiers
urn:nbn:se:umu:diva-95192 (URN)10.1016/j.bpj.2014.09.005 (DOI)000343682700021 ()25418176 (PubMedID)
Available from: 2014-10-23 Created: 2014-10-23 Last updated: 2018-06-07Bibliographically approved
Horvath, I., Sellstedt, M., Weise, C., Nordvall, L.-M., Golla, K. P., Olofsson, A., . . . Wittung-Stafshede, P. (2013). Modulation of α-synuclein fibrillization by ring-fused 2-pyridones: templation and inhibition involve oligomers with different structure. Archives of Biochemistry and Biophysics, 532(2), 84-90
Open this publication in new window or tab >>Modulation of α-synuclein fibrillization by ring-fused 2-pyridones: templation and inhibition involve oligomers with different structure
Show others...
2013 (English)In: Archives of Biochemistry and Biophysics, ISSN 0003-9861, E-ISSN 1096-0384, Vol. 532, no 2, p. 84-90Article in journal (Refereed) Published
Abstract [en]

In a recent study we discovered that a ring-fused 2-pyridone compound triggered fibrillization of a key protein in Parkinson's disease, α-synuclein. To reveal how variations in compound structure affect protein aggregation, we now prepared a number of strategic analogs and tested their effects on α-synuclein amyloid fiber formation in vitro. We find that, in contrast to the earlier templating effect, some analogs inhibit α-synuclein fibrillization. For both templating and inhibiting compounds, the key species formed in the reactions are α-synuclein oligomers that contain compound. Despite similar macroscopic appearance, the templating and inhibiting oligomers are distinctly different in secondary structure content. When the inhibitory oligomers are added in seed amounts, they inhibit fresh α-synuclein aggregation reactions. Our study demonstrates that small chemical changes to the same central fragment can result in opposite effects on protein aggregation.

Place, publisher, year, edition, pages
Elsevier, 2013
Keywords
α-synuclein, amyloid, oligomer, protein aggregation, spectroscopy, 2-pyridone
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-66116 (URN)10.1016/j.abb.2013.01.012 (DOI)000317257700004 ()23399432 (PubMedID)
Available from: 2013-02-14 Created: 2013-02-14 Last updated: 2018-06-08Bibliographically approved
Ådén, J., Weise, C., Brännström, K., Olofsson, A. & Wolf-Watz, M. (2013). Structural topology and activation of an initial adenylate kinase-substrate complex. Biochemistry, 52(6), 1055-1061
Open this publication in new window or tab >>Structural topology and activation of an initial adenylate kinase-substrate complex
Show others...
2013 (English)In: Biochemistry, ISSN 0006-2960, E-ISSN 1520-4995, Vol. 52, no 6, p. 1055-1061Article in journal (Refereed) Published
Abstract [en]

Enzymatic activity is ultimately defined by the structure, chemistry and dynamics of the Michaelis complex. There exist a large number of experimentally determined structures between enzymes and substrates or substrate analogues or inhibitors. However, transient, short-lived encounter and equilibrium structures also play fundamental roles during enzymatic reaction cycles. Such structures are inherently difficult to study with conventional experimental techniques. The enzyme adenylate kinase undergoes major conformational rearrangements in response to binding of its substrates ATP and AMP. ATP is sandwiched between two binding surfaces in the closed and active enzyme conformation. Thus, ade-nylate kinase harbors two spatially distant surfaces in the substrate free open conformation of which one is responsible for the initial interaction with ATP. Here, we have performed primarily nuclear magnetic resonance experiments on Escherichia coli adenylate kinase (AKeco) variants that enabled identification of the site responsible for the initial ATP interaction. This allowed a characterization of the structural topology of an initial equilibrium complex between AKeco and ATP. Based on the results it is suggested that the ATP binding mechanism to AKeco is a mixture between "induced fit" and "conformational selection" models. It is shown that ATP is activated in the initial enzyme bound complex since it displays an appreciable rate of non-productive ATP hydrolysis. In summary our results provide novel structural and functional insights into adenylate kinase catalysis.

National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-64932 (URN)10.1021/bi301460k (DOI)000315018600008 ()23339454 (PubMedID)
Available from: 2013-02-04 Created: 2013-02-04 Last updated: 2018-06-08Bibliographically approved
Frost, S., Ho, O., Login, F. H., Weise, C. F., Wolf-Watz, H. & Wolf-Watz, M. (2012). Autoproteolysis and Intramolecular Dissociation of Yersinia YscU Precedes Secretion of Its C-Terminal Polypeptide YscU CC. PLoS ONE, 7(11), Article ID e49349.
Open this publication in new window or tab >>Autoproteolysis and Intramolecular Dissociation of Yersinia YscU Precedes Secretion of Its C-Terminal Polypeptide YscU CC
Show others...
2012 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 7, no 11, article id e49349Article in journal (Refereed) Published
Abstract [en]

Type III secretion system mediated secretion and translocation of Yop-effector proteins across the eukaryotic target cell membrane by pathogenic Yersinia is highly organized and is dependent on a switching event from secretion of early structural substrates to late effector substrates (Yops). Substrate switching can be mimicked in vitro by modulating the calcium levels in the growth medium. YscU that is essential for regulation of this switch undergoes autoproteolysis at a conserved N↑PTH motif, resulting in a 10 kDa C-terminal polypeptide fragment denoted YscUCC. Here we show that depletion of calcium induces intramolecular dissociation of YscUCC from YscU followed by secretion of the YscUCC polypeptide. Thus, YscUCC behaved in vivo as a Yop protein with respect to secretion properties. Further, destabilized yscU mutants displayed increased rates of dissociation of YscUCC in vitro resulting in enhanced Yop secretion in vivo at 30°C relative to the wild-type strain.These findings provide strong support to the relevance of YscUCC dissociation for Yop secretion. We propose that YscUCC orchestrates a block in the secretion channel that is eliminated by calcium depletion. Further, the striking homology between different members of the YscU/FlhB family suggests that this protein family possess regulatory functions also in other bacteria using comparable mechanisms.

National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-61703 (URN)10.1371/journal.pone.0049349 (DOI)000311821000040 ()23185318 (PubMedID)
Available from: 2012-11-23 Created: 2012-11-23 Last updated: 2018-06-08Bibliographically approved
Niemiec, M. S., Weise, C. F. & Wittung-Stafshede, P. (2012). In vitro thermodynamic dissection of human copper transfer from chaperone to target protein. PLoS ONE, 7(5), e36102
Open this publication in new window or tab >>In vitro thermodynamic dissection of human copper transfer from chaperone to target protein
2012 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 7, no 5, p. e36102-Article in journal (Refereed) Published
Abstract [en]

Transient protein-protein and protein-ligand interactions are fundamental components of biological activity. To understand biological activity, not only the structures of the involved proteins are important but also the energetics of the individual steps of a reaction. Here we use in vitro biophysical methods to deduce thermodynamic parameters of copper (Cu) transfer from the human copper chaperone Atox1 to the fourth metal-binding domain of the Wilson disease protein (WD4). Atox1 and WD4 have the same fold (ferredoxin-like fold) and Cu-binding site (two surface exposed cysteine residues) and thus it is not clear what drives metal transfer from one protein to the other. Cu transfer is a two-step reaction involving a metal-dependent ternary complex in which the metal is coordinated by cysteines from both proteins (i.e., Atox1-Cu-WD4). We employ size exclusion chromatography to estimate individual equilibrium constants for the two steps. This information together with calorimetric titration data are used to reveal enthalpic and entropic contributions of each step in the transfer process. Upon combining the equilibrium constants for both steps, a metal exchange factor (from Atox1 to WD4) of 10 is calculated, governed by a negative net enthalpy change of ∼10 kJ/mol. Thus, small variations in interaction energies, not always obvious upon comparing protein structures alone, may fuel vectorial metal transfer.

National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-56355 (URN)10.1371/journal.pone.0036102 (DOI)000305349800032 ()22574136 (PubMedID)
Available from: 2012-06-14 Created: 2012-06-14 Last updated: 2018-06-08Bibliographically approved
Horvath, I., Weise, C. F., Andersson, E. K., Chorell, E., Sellstedt, M., Bengtsson, C., . . . Wittung-Stafshede, P. (2012). Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation. Journal of the American Chemical Society, 134(7), 3439-3444
Open this publication in new window or tab >>Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
Show others...
2012 (English)In: Journal of the American Chemical Society, ISSN 0002-7863, E-ISSN 1520-5126, Vol. 134, no 7, p. 3439-3444Article in journal (Refereed) Published
Abstract [en]

Small organic molecules that inhibit functional bacterial amyloid fibers, curli, are promising new antibiotics. Here we investigated the mechanism by which the ring-fused 2-pyridone FN075 inhibits fibrillation of the curli protein CsgA. Using a variety of biophysical techniques, we found that FN075 promotes CsgA to form off-pathway, non-amyloidogenic oligomeric species. In light of the generic properties of amyloids, we tested whether FN075 would also affect the fibrillation reaction of human α-synuclein, an amyloid-forming protein involved in Parkinson's disease. Surprisingly, FN075 stimulates α-synuclein amyloid fiber formation as measured by thioflavin T emission, electron microscopy (EM), and atomic force microscopy (AFM). NMR data on (15)N-labeled α-synuclein show that upon FN075 addition, α-synuclein oligomers with 7 nm radius form in which the C-terminal 40 residues remain disordered and solvent exposed. The polypeptides in these oligomers contain β-like secondary structure, and the oligomers are detectable by AFM, EM, and size-exclusion chromatography (SEC). Taken together, FN075 triggers oligomer formation of both proteins: in the case of CsgA, the oligomers do not proceed to fibers, whereas for α-synuclein, the oligomers are poised to rapidly form fibers. We conclude that there is a fine balance between small-molecule inhibition and templation that depends on protein chemistry.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2012
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-52526 (URN)10.1021/ja209829m (DOI)22260746 (PubMedID)
Available from: 2012-02-24 Created: 2012-02-24 Last updated: 2018-06-08Bibliographically approved
Palm, M. E., Weise, C. F., Lundin, C., Wingsle, G., Nygren, Y., Björn, E., . . . Wittung-Stafshede, P. (2011). Cisplatin binds human copper chaperone Atox1 and promotes unfolding in vitro. Proceedings of the National Academy of Sciences of the United States of America, 108(17), 6951-6956
Open this publication in new window or tab >>Cisplatin binds human copper chaperone Atox1 and promotes unfolding in vitro
Show others...
2011 (English)In: Proceedings of the National Academy of Sciences of the United States of America, ISSN 0027-8424, E-ISSN 1091-6490, Vol. 108, no 17, p. 6951-6956Article in journal (Refereed) Published
Abstract [en]

Cisplatin (cisPt), Pt(NH(3))(2)Cl(2), is a cancer drug believed to kill cells via DNA binding and damage. Recent work has implied that the cellular copper (Cu) transport machinery may be involved in cisPt cell export and drug resistance. Normally, the Cu chaperone Atox1 binds Cu(I) via two cysteines and delivers the metal to metal-binding domains of ATP7B; the ATP7B domains then transfer the metal to the Golgi lumen for loading on cuproenzymes. Here, we use spectroscopic methods to test if cisPt interacts with purified Atox1 in solution in vitro. We find that cisPt binds to Atox1's metal-binding site regardless of the presence of Cu or not: When Cu is bound to Atox1, the near-UV circular dichroism signals indicate Cu-Pt interactions. From NMR data, it is evident that cisPt binds to the folded protein. CisPt-bound Atox1 is however not stable over time and the protein begins to unfold and aggregate. The reaction rates are limited by slow cisPt dechlorination. CisPt-induced unfolding of Atox1 is specific because this effect was not observed for two unrelated proteins that also bind cisPt. Our study demonstrates that Atox1 is a candidate for cisPt drug resistance: By binding to Atox1 in the cytoplasm, cisPt transport to DNA may be blocked. In agreement with this model, cell line studies demonstrate a correlation between Atox1 expression levels, and cisplatin resistance.

National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-43722 (URN)10.1073/pnas.1012899108 (DOI)21482801 (PubMedID)
Available from: 2011-05-09 Created: 2011-05-09 Last updated: 2018-06-08Bibliographically approved
Ådén, J., Wallgren, M., Storm, P., Weise, C., Christiansen, A., Schröder, W. P., . . . Wolf-Watz, M. (2011). Extraordinary μs-ms backbone dynamics in Arabidopsis thaliana peroxiredoxin Q. Biochimica et Biophysica Acta, 1814(12), 1880-1890
Open this publication in new window or tab >>Extraordinary μs-ms backbone dynamics in Arabidopsis thaliana peroxiredoxin Q
Show others...
2011 (English)In: Biochimica et Biophysica Acta, ISSN 0006-3002, E-ISSN 1878-2434, Vol. 1814, no 12, p. 1880-1890Article in journal (Refereed) Published
Abstract [en]

Peroxiredoxin Q (PrxQ) isolated from Arabidopsis thaliana belongs to a family of redox enzymes called peroxiredoxins, which are thioredoxin- or glutaredoxin-dependent peroxidases acting to reduce peroxides and in particular hydrogen peroxide. PrxQ cycles between an active reduced state and an inactive oxidized state during its catalytic cycle. The catalytic mechanism involves a nucleophilic attack of the catalytic cysteine on hydrogen peroxide to generate a sulfonic acid intermediate with a concerted release of a water molecule. This intermediate is subsequently relaxed by the reaction of a second cysteine, denoted the resolving cysteine, generating an intramolecular disulfide bond and release of a second water molecule. PrxQ is recycled to the active state by a thioredoxin-dependent reduction. Previous structural studies of PrxQ homologues have provided the structural basis for the switch between reduced and oxidized conformations. Here, we have performed a detailed study of the activity, structure and dynamics of PrxQ in both the oxidized and reduced states. Reliable and experimentally validated structural models of PrxQ in both oxidation states were generated using homology based modeling. Analysis of NMR spin relaxation rates shows that PrxQ is monomeric in both oxidized and reduced states. As evident from R(2) relaxation rates the reduced form of PrxQ undergoes unprecedented dynamics on the slow μs-ms timescale. The ground state of this conformational dynamics is likely the stably folded reduced state as implied by circular dichroism spectroscopy. We speculate that the extensive dynamics is intimately related to the catalytic function of PrxQ.

Place, publisher, year, edition, pages
Elsevier, 2011
Keywords
NMR, Enzyme, Dynamics, Peroxiredoxin, Arabidopsis thaliana
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-45749 (URN)10.1016/j.bbapap.2011.07.011 (DOI)21798375 (PubMedID)
Note
Received 1 April 2011; revised 27 June 2011; Accepted 12 July 2011. Available online 21 July 2011. In Press, Corrected Proof - Note to usersAvailable from: 2011-08-16 Created: 2011-08-16 Last updated: 2018-06-08Bibliographically approved
Aguilar, X., Weise, C., Sparrman, T., Wolf-Watz, M. & Wittung-Stafshede, P. (2011). Macromolecular crowding extended to a heptameric system: the co-chaperonin protein 10. Biochemistry, 50(14), 3034-3044
Open this publication in new window or tab >>Macromolecular crowding extended to a heptameric system: the co-chaperonin protein 10
Show others...
2011 (English)In: Biochemistry, ISSN 0006-2960, E-ISSN 1520-4995, Vol. 50, no 14, p. 3034-3044Article in journal (Refereed) Published
Abstract [en]

Experiments on monomeric proteins have shown that macromolecular crowding can stabilize toward heat perturbation and also modulate native-state structure. To assess the effects of macromolecular crowding on unfolding of an oligomeric protein, we here tested the effects of the synthetic crowding agent Ficoll 70 on human cpn10 (GroES in E. coli), a heptameric protein consisting of seven identical β-barrel subunits assembling into a ring. Using far-UV circular dichroism (CD), tyrosine fluorescence, nuclear magnetic resonance (NMR), and cross-linking experiments, we investigated thermal and chemical stability, as well as the heptamer-monomer dissociation constant, without and with crowding agent. We find that crowding shifts the heptamer-monomer equilibrium constant in the direction of the heptamer. The cpn10 heptamer is both thermally and thermodynamically stabilized in 300 mg/mL Ficoll 70 as compared to regular buffer conditions. Kinetic unfolding experiments show that the increased stability in crowded conditions, in part, is explained by slower unfolding rates. A thermodynamic cycle reveals that in presence of 300 mg/mL Ficoll the thermodynamic stability of each cpn10 monomer increases by over 30%, whereas the interfaces are stabilized by less than 10%. We also introduce a new approach to analyze the spectroscopic data that makes use of multiple wavelengths: this provides robust error estimates of thermodynamic parameters.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2011
National Category
Chemical Sciences
Identifiers
urn:nbn:se:umu:diva-43730 (URN)10.1021/bi2002086 (DOI)21375247 (PubMedID)
Available from: 2011-05-09 Created: 2011-05-09 Last updated: 2018-06-08Bibliographically approved
Organisations

Search in DiVA

Show all publications