• Login
    View Item 
    •   Home
    • Research
    • Articles
    • View Item
    •   Home
    • Research
    • Articles
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of KAUSTCommunitiesIssue DateSubmit DateThis CollectionIssue DateSubmit Date

    My Account

    Login

    Quick Links

    Open Access PolicyORCID LibguidePlumX LibguideSubmit an Item

    Statistics

    Display statistics

    Biochemical Stability and Molecular Dynamic Characterization of Aspergillus fumigatus Cystathionine γ-Lyase in Response to Various Reaction Effectors

    • CSV
    • RefMan
    • EndNote
    • BibTex
    • RefWorks
    Thumbnail
    Name:
    1-s2.0-S0141022915300399-main.pdf
    Size:
    7.587Mb
    Format:
    PDF
    Description:
    Accepted Manuscript
    Download
    Thumbnail
    Name:
    mmc1.docx
    Size:
    1.180Mb
    Format:
    Microsoft Word 2007
    Description:
    Supplemental files
    Download
    Type
    Article
    Authors
    El-Sayed, Ashraf S.A.
    Abdel-Azeim, Safwat cc
    Ibrahim, Hend M.
    Yassin, Marwa A.
    Abdel-Ghany, Salah E.
    Esener, Sadik
    Ali, Gul Shad
    KAUST Department
    KAUST Catalysis Center (KCC)
    Date
    2015-08-11
    Online Publication Date
    2015-08-11
    Print Publication Date
    2015-12
    Permanent link to this record
    http://hdl.handle.net/10754/567038
    
    Metadata
    Show full item record
    Abstract
    Cystathionine γ-lyase (CGL) is a key enzyme in the methionine-cysteine cycle in all living organisms forming cysteine, α-ketobutyrate and ammonia via homocysteine and cystathionine intermediates. Although, human and plant CGLs have been extensively studied at the molecular and mechanistic levels, there has been little work on the molecular and catalytic properties of fungal CGL. Herein, we studied in detail for the first time the molecular and catalytic stability of Aspergillus fumigatus CGL, since conformational instability, inactivation and structural antigenicity are the main limitations of the PLP-dependent enzymes on various therapeutic uses. We examined these properties in response to buffer compositions, stabilizing and destabilizing agents using Differential Scanning Fluorometery (DSF), steady state and gel-based fluorescence of the intrinsic hydrophobic core, stability of internal aldimine linkage and catalytic properties. The activity of the recombinant A. fumigatus CGL was 13.8 U/mg. The melting temperature (Tm) of CGL in potassium phosphate buffer (pH 7.0-8.0) was 73.3 °C, with ∼3 °C upshifting in MES and sodium phosphate buffers (pH 7.0). The conformational thermal stability was increased in potassium phosphate, sodium phosphate and MES buffers, in contrast to Tris-HCl, HEPES (pH 7.0) and CAPS (pH 9.0-10.0). The thermal stability and activity of CGL was slightly increased in the presence of trehalose and glycerol that might be due to hydration of the enzyme backbone, unlike the denaturing effect of GdmCl and urea. Modification of surface CGL glutamic and aspartic acids had no significant effect on the enzyme conformational and catalytic stability. Molecular modeling and dynamics simulations unveil the high conformational stability of the overall scaffold of CGL with high flexibility at the non-structural regions. CGL structure has eight buried Trp residues, which are reoriented to the enzyme surface and get exposed to the solvent under perturbation of destabilizers. Furthermore, electrostatic calculations of selected snapshots of CGL 3D structure under different experimental conditions showed a remarkable differences on the polarity of the enzyme surface.
    Citation
    Biochemical Stability and Molecular Dynamic Characterization of Aspergillus fumigatus Cystathionine γ-Lyase in Response to Various Reaction Effectors 2015 Enzyme and Microbial Technology
    Publisher
    Elsevier BV
    Journal
    Enzyme and Microbial Technology
    DOI
    10.1016/j.enzmictec.2015.08.004
    PubMed ID
    26453470
    Additional Links
    http://linkinghub.elsevier.com/retrieve/pii/S0141022915300399
    ae974a485f413a2113503eed53cd6c53
    10.1016/j.enzmictec.2015.08.004
    Scopus Count
    Collections
    Articles; KAUST Catalysis Center (KCC)

    entitlement

    Related articles

    • Biochemical and Pharmacokinetic Properties of PEGylated Cystathionine γ-Lyase from Aspergillus carneus KF723837.
    • Authors: El-Sayed AS, Yassin MA, Khalaf SA, El-Batrik M, Ali GS, Esener S
    • Issue date: 2015
    • A role for glutamate-333 of Saccharomyces cerevisiae cystathionine γ-lyase as a determinant of specificity.
    • Authors: Hopwood EM, Ahmed D, Aitken SM
    • Issue date: 2014 Feb
    • Determinants of enzymatic specificity in the Cys-Met-metabolism PLP-dependent enzymes family: crystal structure of cystathionine gamma-lyase from yeast and intrafamiliar structure comparison.
    • Authors: Messerschmidt A, Worbs M, Steegborn C, Wahl MC, Huber R, Laber B, Clausen T
    • Issue date: 2003 Mar
    • Structural Snapshots of an Engineered Cystathionine-γ-lyase Reveal the Critical Role of Electrostatic Interactions in the Active Site.
    • Authors: Yan W, Stone E, Zhang YJ
    • Issue date: 2017 Feb 14
    • Molecular and Spectroscopic Characterization of Aspergillus flavipes and Pseudomonas putida L-Methionine γ-Lyase in Vitro.
    • Authors: El-Sayed AS, Ruff LE, Ghany SE, Ali GS, Esener S
    • Issue date: 2017 Apr
    DSpace software copyright © 2002-2021  DuraSpace
    Quick Guide | Contact Us | Send Feedback
    Open Repository is a service hosted by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items. For anonymous users the allowed maximum amount is 50 search results.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.