ACS Publications. Most Trusted. Most Cited. Most Read
Cation Binding and Thermostability of FTHFS Monovalent Cation Binding Sites and Thermostability of N10-Formyltetrahydrofolate Synthetase from Moorella thermoacetica,

OR SEARCH CITATIONS

My Activity
Publications

Figure 1Loading Img
Download Hi-Res ImageDownload to MS-PowerPointCite This:Biochemistry 2000, 39, 47, 14481-14486
    Article

    Cation Binding and Thermostability of FTHFS Monovalent Cation Binding Sites and Thermostability of N10-Formyltetrahydrofolate Synthetase from Moorella thermoacetica†,‡
    Click to copy article linkArticle link copied!

    View Author Information
    Departments of Chemistry and Biochemistry and Biological Sciences, University of South Carolina, Columbia, South Carolina 29208, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602, and Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22901
    Other Access Options

    Biochemistry

    Cite this: Biochemistry 2000, 39, 47, 14481–14486
    Click to copy citationCitation copied!
    https://doi.org/10.1021/bi001577w
    Published October 26, 2000
    Copyright © 2000 American Chemical Society
    Request reuse permissions

    Abstract

    Click to copy section linkSection link copied!

    Formyltetrahydrofolate synthetase (FTHFS) from the thermophilic homoacetogen, Moorella thermoacetica, has an optimum temperature for activity of 55−60 °C and requires monovalent cations for both optimal activity and stabilization of tetrameric structure at higher temperatures. The crystal structures of complexes of FTHFS with cesium and potassium ions were examined and monovalent cation binding positions identified. Unexpectedly, NH4+ and K+, both of which are strongly activating ions, bind at a different site than a moderately activating ion, Cs+, does. Neither binding site is located in the active site. The sites are 7 Å apart, but in each of them, the side chain of Glu 98, which is conserved in all known bacterial FTHFS sequences, participates in metal ion binding. Other ligands in the Cs+ binding site are four oxygen atoms of main chain carbonyls and water molecules. The K+ and NH4+ binding site includes the carboxylate of Asp132 in addition to Glu98. Mutant FTHFS's (E98Q, E98D, and E98S) were obtained and analyzed using differential scanning calorimetry to examine the effect of these mutations on the thermostability of the enzyme with and without added K+ ions. The addition of 0.2 M K+ ions to the wild-type enzyme resulted in a 10 °C increase in the thermal denaturation temperature. No significant increase was observed in E98D or E98S. The lack of a significant effect of monovalent cations on the stability of E98D and E98S indicates that this alteration of the binding site eliminates cation binding. The thermal denaturation temperature of E98Q was 3 °C higher than that of the wild-type enzyme in the absence of the cation, indicating that the removal of the unbalanced, buried charge of Glu98 stabilizes the enzyme. These results confirm that Glu98 is a crucial residue in the interaction of monovalent cations with FTHFS.

    Copyright © 2000 American Chemical Society

    Subjects

    what are subjects

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.

    Recommended

    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. Add or change your institution or let them know you’d like them to include access.

    †

     This work was supported by NSF Grant MCB-9873606. Some instrumentation used in this research was purchased with NSF Grant BIR 9419866 and DOE Grant DE-FG-95TE00058. Use of the Argonne National Laboratory Structural Biology Center beamlines at the Advanced Photon Source was supported by the U.S. Department of Energy, Office of Science, under Contract W-31-109-ENG-38.

    ‡

     The PDB files of the atomic coordinates of Cs+ and K+ complexes of N10-formyltetrahydrofolate synthetase are deposited in the Protein Data Bank as entries 1fpm and 1fp7.

    §

     Department of Chemistry and Biochemistry, University of South Carolina.

    ‖

     These authors contributed equally to the described research.

    ⊥

     Department of Biological Sciences, University of South Carolina.

    @

     University of Georgia.

    #

     University of Virginia.

    *

     To whom correspondence should be addressed:  Department of Chemistry and Biochemistry, University of South Carolina, 631 Sumter St., Columbia, SC 29208. E-mail:  [email protected]. Phone:  (803) 777-2140. Fax:  (803) 777-9521.

    Cited By

    Click to copy section linkSection link copied!
    Citation Statements
    Explore this article's citation statements on scite.ai
    powered by  Scite

    This article is cited by 17 publications.

    1. Kamyll Dawn Cocon, Patricia Luis. The potential of RuBisCO in CO2 capture and utilization. Progress in Energy and Combustion Science 2024, 105 , 101184. https://doi.org/10.1016/j.pecs.2024.101184
    2. Elahe Abedi, Shima Kaveh, Seyed Mohammad Bagher Hashemi. Structure-based modification of a-amylase by conventional and emerging technologies: Comparative study on the secondary structure, activity, thermal stability and amylolysis efficiency. Food Chemistry 2024, 437 , 137903. https://doi.org/10.1016/j.foodchem.2023.137903
    3. Kyeong Rok Choi, Yeah-Ji Ahn, Sang Yup Lee. Bacterial conversion of CO2 to organic compounds. Journal of CO2 Utilization 2022, 58 , 101929. https://doi.org/10.1016/j.jcou.2022.101929
    4. Long-Liu Lin, Bo-Yuan Lu, Meng-Chun Chi, Yu-Fen Huang, Min-Guan Lin, Tzu-Fan Wang. Activation and thermal stabilization of a recombinant γ-glutamyltranspeptidase from Bacillus licheniformis ATCC 27811 by monovalent cations. Applied Microbiology and Biotechnology 2022, 106 (5-6) , 1991-2006. https://doi.org/10.1007/s00253-022-11836-y
    5. Jimyung Moon, Judith Dönig, Sina Kramer, Anja Poehlein, Rolf Daniel, Volker Müller. Formate metabolism in the acetogenic bacterium Acetobacterium woodii. Environmental Microbiology 2021, 23 (8) , 4214-4227. https://doi.org/10.1111/1462-2920.15598
    6. Boguslaw Stec. Time passes yet errors remain: Comments on the structure of N 10 ‐formyltetrahydrofolate synthetase. Protein Science 2013, 22 (6) , 671-674. https://doi.org/10.1002/pro.2252
    7. Bettina Müller, Li Sun, Anna Schnürer. First insights into the syntrophic acetate‐oxidizing bacteria – a genetic study. MicrobiologyOpen 2013, 2 (1) , 35-53. https://doi.org/10.1002/mbo3.50
    8. Lesa R. Celeste, Geqing Chai, Magdalena Bielak, Wladek Minor, Leslie L. Lovelace, Lukasz Lebioda. Mechanism of N 10 ‐formyltetrahydrofolate synthetase derived from complexes with intermediates and inhibitors. Protein Science 2012, 21 (2) , 219-228. https://doi.org/10.1002/pro.2005
    9. Gemma Henderson, Graham E. Naylor, Sinead C. Leahy, Peter H. Janssen. Presence of Novel, Potentially Homoacetogenic Bacteria in the Rumen as Determined by Analysis of Formyltetrahydrofolate Synthetase Sequences from Ruminants. Applied and Environmental Microbiology 2010, 76 (7) , 2058-2066. https://doi.org/10.1128/AEM.02580-09
    10. W.F. Li, X.X. Zhou, P. Lu. Structural features of thermozymes. Biotechnology Advances 2005, 23 (4) , 271-281. https://doi.org/10.1016/j.biotechadv.2005.01.002
    11. Satoshi Hattori, Alexander S. Galushko, Yoichi Kamagata, Bernhard Schink. Operation of the CO Dehydrogenase/Acetyl Coenzyme A Pathway in both Acetate Oxidation and Acetate Formation by the Syntrophically Acetate-Oxidizing Bacterium Thermacetogenium phaeum. Journal of Bacteriology 2005, 187 (10) , 3471-3476. https://doi.org/10.1128/JB.187.10.3471-3476.2005
    12. Swati Prasad, Kelly J. Wright, Dolly Banerjee Roy, Leslie A. Bush, Angelene M. Cantwell, Enrico Di Cera. Redesigning the monovalent cation specificity of an enzyme. Proceedings of the National Academy of Sciences 2003, 100 (24) , 13785-13790. https://doi.org/10.1073/pnas.2333109100
    13. Tina M. Salmassi, Jared R. Leadbetter. Analysis of genes of tetrahydrofolate-dependent metabolism from cultivated spirochaetes and the gut community of the termite Zootermopsis angusticollis. Microbiology 2003, 149 (9) , 2529-2537. https://doi.org/10.1099/mic.0.26351-0
    14. Jaeho Lee, Sergiu M. Gorun. Compositional and Isotopologue‐Induced Phase Differentiation in Supramolecular Aggregates. Angewandte Chemie 2003, 115 (13) , 1550-1553. https://doi.org/10.1002/ange.200250200
    15. Jaeho Lee, Sergiu M. Gorun. Compositional and Isotopologue‐Induced Phase Differentiation in Supramolecular Aggregates. Angewandte Chemie International Edition 2003, 42 (13) , 1512-1515. https://doi.org/10.1002/anie.200250200
    16. Adam B Leaphart, H Trent Spencer, Charles R Lovell. Site-directed mutagenesis of a potential catalytic and formyl phosphate binding site and substrate inhibition of N10-formyltetrahydrofolate synthetase. Archives of Biochemistry and Biophysics 2002, 408 (1) , 137-143. https://doi.org/10.1016/S0003-9861(02)00552-0
    17. C.Evalena Andersson, Sherry L Mowbray. Activation of ribokinase by monovalent cations. Journal of Molecular Biology 2002, 315 (3) , 409-419. https://doi.org/10.1006/jmbi.2001.5248
    Download PDF
    Go to Biochemistry
    Get e-Alerts
    Get e-Alerts

    Biochemistry

    Cite this: Biochemistry 2000, 39, 47, 14481–14486
    Click to copy citationCitation copied!
    https://doi.org/10.1021/bi001577w
    Published October 26, 2000
    Copyright © 2000 American Chemical Society
    Request reuse permissions

    Article Views

    462

    Altmetric

    -

    Citations

    17
    Learn about these metrics

    Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

    Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

    The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.