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Hainzl, Tobias
Publications (8 of 8) Show all publications
Benlloch, R., Shevela, D., Hainzl, T., Grundström, C., Shutova, T., Messinger, J., . . . Sauer-Eriksson, E. (2015). Crystal structure and functional characterization of Photosystem II-associated carbonic anhydrase CAH3 in Chlamydomonas reinhardtii. Plant Physiology, 167(3), 950-962
Open this publication in new window or tab >>Crystal structure and functional characterization of Photosystem II-associated carbonic anhydrase CAH3 in Chlamydomonas reinhardtii
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2015 (English)In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 167, no 3, p. 950-962Article in journal (Refereed) Published
Abstract [en]

In oxygenic photosynthesis, light energy is stored in the form of chemical energy by converting CO2 and water into carbohydrates.The light-driven oxidation of water that provides the electrons and protons for the subsequent CO2 fixation takes place inphotosystem II (PSII). Recent studies show that in higher plants, HCO3– increases PSII activity by acting as a mobile acceptor ofthe protons produced by PSII. In the green alga Chlamydomonas reinhardtii, a luminal carbonic anhydrase, CrCAH3, was suggested toimprove proton removal from PSII, possibly by rapid reformation of HCO3– from CO2. In this study, we investigated the interplaybetween PSII and CrCAH3 by membrane inlet mass spectrometry and x-ray crystallography. Membrane inlet mass spectrometrymeasurements showed that CrCAH3 was most active at the slightly acidic pH values prevalent in the thylakoid lumen underillumination. Two crystal structures of CrCAH3 in complex with either acetazolamide or phosphate ions were determined at 2.6- and2.7-Å resolution, respectively. CrCAH3 is a dimer at pH 4.1 that is stabilized by swapping of the N-terminal arms, a feature notpreviously observed in a-type carbonic anhydrases. The structure contains a disulfide bond, and redox titration of CrCAH3 functionwith dithiothreitol suggested a possible redox regulation of the enzyme. The stimulating effect of CrCAH3 and CO2/HCO3– on PSIIactivity was demonstrated by comparing the flash-induced oxygen evolution pattern of wild-type and CrCAH3-less PSIIpreparations. We showed that CrCAH3 has unique structural features that allow this enzyme to maximize PSII activity at lowpH and CO2 concentration.

Place, publisher, year, edition, pages
American Society of Plant Biologists, 2015
National Category
Botany Biochemistry and Molecular Biology
Research subject
biological chemistry; Biochemistry
Identifiers
urn:nbn:se:umu:diva-103651 (URN)10.1104/pp.114.253591 (DOI)000354413900027 ()
Funder
Knut and Alice Wallenberg FoundationSwedish Research Council
Available from: 2015-05-26 Created: 2015-05-26 Last updated: 2018-06-07Bibliographically approved
Hainzl, T. & Sauer-Eriksson, A. E. (2015). Signal-sequence induced conformational changes in the signal recognition particle. Nature Communications, 6, Article ID 7163.
Open this publication in new window or tab >>Signal-sequence induced conformational changes in the signal recognition particle
2015 (English)In: Nature Communications, ISSN 2041-1723, E-ISSN 2041-1723, Vol. 6, article id 7163Article in journal (Refereed) Published
Abstract [en]

Co-translational protein targeting is an essential, evolutionarily conserved pathway for delivering nascent proteins to the proper cellular membrane. In this pathway, the signal recognition particle (SRP) first recognizes the N-terminal signal sequence of nascent proteins and subsequently interacts with the SRP receptor. For this, signal sequence binding in the SRP54 M domain must be effectively communicated to the SRP54 NG domain that interacts with the receptor. Here we present the 2.9 angstrom crystal structure of unbound- and signal sequence bound SRP forms, both present in the asymmetric unit. The structures provide evidence for a coupled binding and folding mechanism in which signal sequence binding induces the concerted folding of the GM linker helix, the finger loop, and the C-terminal alpha helix alpha M6. This mechanism allows for a high degree of structural adaptability of the binding site and suggests how signal sequence binding in the M domain is coupled to repositioning of the NG domain.

National Category
Organic Chemistry
Identifiers
urn:nbn:se:umu:diva-106604 (URN)10.1038/ncomms8163 (DOI)000357166200002 ()26051119 (PubMedID)
Available from: 2015-07-28 Created: 2015-07-24 Last updated: 2018-06-07Bibliographically approved
Hainzl, T., Huang, S., Meriläinen, G., Brännström, K. & Sauer-Eriksson, A. E. (2011). Structural basis of signal-sequence recognition by the signal recognition particle . Nature Structural & Molecular Biology, 18(3), 389-391
Open this publication in new window or tab >>Structural basis of signal-sequence recognition by the signal recognition particle 
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2011 (English)In: Nature Structural & Molecular Biology, ISSN 1545-9993, E-ISSN 1545-9985, Vol. 18, no 3, p. 389-391Article in journal (Refereed) Published
Abstract [en]

The signal recognition particle (SRP) recognizes and binds the signal sequence of nascent proteins as they emerge from the ribosome. We present here the 3.0-Å structure of a signal sequence bound to the Methanococcus jannaschii SRP core. Structural comparison with the free SRP core shows that signal-sequence binding induces formation of the GM-linker helix and a 180° flip of the NG domain—structural changes that ensure a hierarchical succession of events during protein targeting.

Identifiers
urn:nbn:se:umu:diva-40342 (URN)10.1038/nsmb.1994 (DOI)
Note
Published online 20 February 2011Available from: 2011-02-22 Created: 2011-02-22 Last updated: 2018-06-08Bibliographically approved
Huang, S., Hainzl, T., Grundström, C., Forsman, C., Samuelsson, G. & Sauer-Eriksson, A. E. (2011). Structural studies of β-Carbonic Anhydrase from the Green Alga Coccomyxa: Inhibitor complexes with Anions and Acetazolamide. PLoS ONE, 6(12), e28458
Open this publication in new window or tab >>Structural studies of β-Carbonic Anhydrase from the Green Alga Coccomyxa: Inhibitor complexes with Anions and Acetazolamide
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2011 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 6, no 12, p. e28458-Article in journal (Refereed) Published
Abstract [en]

The β-class carbonic anhydrases (β-CAs) are widely distributed among lower eukaryotes, prokaryotes, archaea, and plants. Like all CAs, the β-enzymes catalyze an important physiological reaction, namely the interconversion between carbon dioxide and bicarbonate. In plants the enzyme plays an important role in carbon fixation and metabolism. To further explore the structure-function relationship of β-CA, we have determined the crystal structures of the photoautotroph unicellular green alga Coccomyxa β-CA in complex with five different inhibitors: acetazolamide, thiocyanate, azide, iodide, and phosphate ions. The tetrameric Coccomyxa β-CA structure is similar to other β-CAs but it has a 15 amino acid extension in the C-terminal end, which stabilizes the tetramer by strengthening the interface. Four of the five inhibitors bind in a manner similar to what is found in complexes with α-type CAs. Iodide ions, however, make contact to the zinc ion via a zinc-bound water molecule or hydroxide ion - a type of binding mode not previously observed in any CA. Binding of inhibitors to Coccomyxa β-CA is mediated by side-chain movements of the conserved residue Tyr-88, extending the width of the active site cavity with 1.5-1.8 Å. Structural analysis and comparisons with other α- and β-class members suggest a catalytic mechanism in which the movements of Tyr-88 are important for the CO(2)-HCO(3) (-) interconversion, whereas a structurally conserved water molecule that bridges residues Tyr-88 and Gln-38, seems important for proton transfer, linking water molecules from the zinc-bound water to His-92 and buffer molecules.

National Category
Structural Biology Biochemistry and Molecular Biology
Research subject
Biochemistry
Identifiers
urn:nbn:se:umu:diva-50781 (URN)10.1371/journal.pone.0028458 (DOI)000298172800038 ()22162771 (PubMedID)
Available from: 2011-12-21 Created: 2011-12-21 Last updated: 2018-06-08Bibliographically approved
Hainzl, T., Huang, S. & Sauer-Eriksson, E. (2007). Interaction of signal-recognition particle 54 GTPase domain and signal-recognition particle RNA in the free signal-recognition particle.. Proc Natl Acad Sci U S A, 104(38), 14911-6
Open this publication in new window or tab >>Interaction of signal-recognition particle 54 GTPase domain and signal-recognition particle RNA in the free signal-recognition particle.
2007 (English)In: Proc Natl Acad Sci U S A, ISSN 0027-8424, Vol. 104, no 38, p. 14911-6Article in journal (Refereed) Published
Abstract [en]

The signal-recognition particle (SRP) is a ubiquitous protein-RNA complex that targets proteins to cellular membranes for insertion or secretion. A key player in SRP-mediated protein targeting is the evolutionarily conserved core consisting of the SRP RNA and the multidomain protein SRP54. Communication between the SRP54 domains is critical for SRP function, where signal sequence binding at the M domain directs receptor binding at the GTPase domain (NG domain). These SRP activities are linked to domain rearrangements, for which the role of SRP RNA is not clear. In free SRP, a direct interaction of the GTPase domain with SRP RNA has been proposed but has never been structurally verified. In this study, we present the crystal structure at 2.5-A resolution of the SRP54-SRP19-SRP RNA complex of Methanococcus jannaschii SRP. The structure reveals an RNA-bound conformation of the SRP54 GTPase domain, in which the domain is spatially well separated from the signal peptide binding site. The association of both the N and G domains with SRP RNA in free SRP provides further structural evidence for the pivotal role of SRP RNA in the regulation of the SRP54 activity.

Identifiers
urn:nbn:se:umu:diva-16829 (URN)17846429 (PubMedID)
Available from: 2007-10-12 Created: 2007-10-12 Last updated: 2018-06-09Bibliographically approved
Hainzl, T., Huang, S. & Sauer-Eriksson, E. (2005). Structural insights into SRP RNA: an induced fit mechanism for SRP assembly.. RNA, 11(7), 1043-50
Open this publication in new window or tab >>Structural insights into SRP RNA: an induced fit mechanism for SRP assembly.
2005 (English)In: RNA, ISSN 1355-8382, Vol. 11, no 7, p. 1043-50Article in journal (Refereed) Published
Abstract [en]

Proper assembly of large protein-RNA complexes requires sequential binding of the proteins to the RNA. The signal recognition particle (SRP) is a multiprotein-RNA complex responsible for the cotranslational targeting of proteins to biological membranes. Here we describe the crystal structure at 2.6-A resolution of the S-domain of SRP RNA from the archeon Methanococcus jannaschii. Comparison of this structure with the SRP19-bound form reveals the nature of the SRP19-induced conformational changes, which promote subsequent SRP54 attachment. These structural changes are initiated at the SRP19 binding site and transmitted through helix 6 to looped-out adenosines, which form tertiary RNA interaction with helix 8. Displacement of these adenosines enforces a conformational change of the asymmetric loop structure in helix 8. In free RNA, the three unpaired bases A195, C196, and C197 are directed toward the helical axis, whereas upon SRP19 binding the loop backbone inverts and the bases are splayed out in a conformation that resembles the SRP54-bound form. Nucleotides adjacent to the bulged nucleotides seem to be particularly important in the regulation of this loop transition. Binding of SRP19 to 7S RNA reveals an elegant mechanism of how protein-induced changes are directed through an RNA molecule and may relate to those regulating the assembly of other RNPs.

Keywords
Adenosine/chemistry, Base Sequence, Binding Sites, Chromatography; Gel, Crystallography; X-Ray, Cytosine/chemistry, Electrophoretic Mobility Shift Assay, Lactococcus lactis/genetics, Methanococcus/*chemistry/genetics, Models; Biological, Models; Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Protein Binding, Protein Structure; Secondary, Protein Structure; Tertiary, RNA; Archaeal/*chemistry/genetics/*metabolism, Signal Recognition Particle/*chemistry/*metabolism
Identifiers
urn:nbn:se:umu:diva-13611 (URN)15928341 (PubMedID)
Available from: 2007-10-12 Created: 2007-10-12 Last updated: 2018-06-09Bibliographically approved
Sauer-Eriksson, E. & Hainzl, T. (2003). S-domain assembly of the signal recognition particle.. Curr Opin Struct Biol, 13(1), 64-70
Open this publication in new window or tab >>S-domain assembly of the signal recognition particle.
2003 (English)In: Curr Opin Struct Biol, ISSN 0959-440X, Vol. 13, no 1, p. 64-70Article, review/survey (Other (popular science, discussion, etc.)) Published
Abstract [en]

The signal recognition particle (SRP) is a phylogenetically conserved ribonucleoprotein that associates with ribosomes to mediate the targeting of membrane and secretory proteins to biological membranes. In higher eukaryotes, SRP biogenesis involves the sequential binding of SRP19 and SRP54 proteins to the S domain of 7S RNA. The recently determined crystal structures of SRP19 in complex with the S domain, and that of the ternary complex of SRP19, the S domain and the M domain of SRP54, provide insight into the molecular basis of S-domain assembly and SRP function.

Keywords
Binding Sites, Crystallography/methods, Escherichia coli/chemistry/metabolism, Humans, Macromolecular Substances, Models; Molecular, Protein Binding, Protein Conformation, Protein Structure; Secondary, Protein Structure; Tertiary, RNA; Small Cytoplasmic/chemistry/metabolism, Signal Recognition Particle/*chemistry/classification/metabolism, Species Specificity, Structure-Activity Relationship
Identifiers
urn:nbn:se:umu:diva-13954 (URN)12581661 (PubMedID)
Available from: 2007-10-12 Created: 2007-10-12 Last updated: 2018-06-09Bibliographically approved
Hainzl, T., Huang, S. & Sauer-Eriksson, E. (2002). Structure of the SRP19 RNA complex and implications for signal recognition particle assembly.. Nature, 417(6890), 767-71
Open this publication in new window or tab >>Structure of the SRP19 RNA complex and implications for signal recognition particle assembly.
2002 (English)In: Nature, ISSN 0028-0836, Vol. 417, no 6890, p. 767-71Article in journal (Refereed) Published
Abstract [en]

The signal recognition particle (SRP) is a phylogenetically conserved ribonucleoprotein. It associates with ribosomes to mediate co-translational targeting of membrane and secretory proteins to biological membranes. In mammalian cells, the SRP consists of a 7S RNA and six protein components. The S domain of SRP comprises the 7S.S part of RNA bound to SRP19, SRP54 and the SRP68/72 heterodimer; SRP54 has the main role in recognizing signal sequences of nascent polypeptide chains and docking SRP to its receptor. During assembly of the SRP, binding of SRP19 precedes and promotes the association of SRP54 (refs 4, 5). Here we report the crystal structure at 2.3 A resolution of the complex formed between 7S.S RNA and SRP19 in the archaeon Methanococcus jannaschii. SRP19 bridges the tips of helices 6 and 8 of 7S.S RNA by forming an extensive network of direct protein RNA interactions. Helices 6 and 8 pack side by side; tertiary RNA interactions, which also involve the strictly conserved tetraloop bases, stabilize helix 8 in a conformation competent for SRP54 binding. The structure explains the role of SRP19 and provides a molecular framework for SRP54 binding and SRP assembly in Eukarya and Archaea.

Keywords
Amino Acid Sequence, Archaeal Proteins/chemistry/metabolism, Base Sequence, Binding Sites, Crystallography; X-Ray, Eukaryotic Cells/chemistry/metabolism, Humans, Methanococcus/chemistry/genetics, Models; Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Protein Conformation, RNA; Archaeal/*chemistry/genetics/*metabolism, Signal Recognition Particle/*chemistry/genetics/*metabolism
Identifiers
urn:nbn:se:umu:diva-13958 (URN)12050674 (PubMedID)
Available from: 2007-10-12 Created: 2007-10-12 Last updated: 2018-06-09Bibliographically approved
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