دورية أكاديمية
In Vitro and In Silico Explorations of the Protein Conformational Changes of Corynebacterium glutamicum MshA, a Model Retaining GT-B Glycosyltransferase.
| العنوان: | In Vitro and In Silico Explorations of the Protein Conformational Changes of Corynebacterium glutamicum MshA, a Model Retaining GT-B Glycosyltransferase. |
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| المؤلفون: | Hassan BA; Department of Chemistry, Wesleyan University, Middletown, Connecticut 06459, United States., Milicaj J; Department of Chemistry, Wesleyan University, Middletown, Connecticut 06459, United States., Tyson M; Department of Chemistry, Wesleyan University, Middletown, Connecticut 06459, United States., Karki R; Department of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama 35487, United States., Sham YY; Bioinformatics and Computational Biology Program, University of Minnesota, Minneapolis, Minnesota 55455, United States.; Department of Integrative Biology and Physiology, Medical School, University of Minnesota, Minneapolis, Minnesota 55455, United States., Frantom PA; Department of Chemistry & Biochemistry, The University of Alabama, Tuscaloosa, Alabama 35487, United States., Taylor EA; Department of Chemistry, Wesleyan University, Middletown, Connecticut 06459, United States. |
| المصدر: | Biochemistry [Biochemistry] 2024 Apr 02; Vol. 63 (7), pp. 939-951. Date of Electronic Publication: 2024 Mar 20. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370623 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-4995 (Electronic) Linking ISSN: 00062960 NLM ISO Abbreviation: Biochemistry Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: | Glycosyltransferases*/metabolism , Corynebacterium glutamicum* , Cysteine* , Glycopeptides* , Inositol*, Ligands ; Inositol Phosphates/metabolism ; Uridine Diphosphate/metabolism ; Protein Conformation ; Molecular Dynamics Simulation |
| مستخلص: | MshA is a GT-B glycosyltransferase catalyzing the first step in the biosynthesis of mycothiol. While many GT-B enzymes undergo an open-to-closed transition, MshA is unique because its 97° rotation is beyond the usual range of 10-25°. Molecular dynamics (MD) simulations were carried out for MshA in both ligand bound and unbound states to investigate the effect of ligand binding on localized protein dynamics and its conformational free energy landscape. Simulations showed that both the unliganded "opened" and liganded "closed" forms of the enzyme sample a wide degree of dihedral angles and interdomain distances with relatively low overlapping populations. Calculation of the free energy surface using replica exchange MD for the apo "opened" and an artificial generated apo "closed" structure revealed overlaps in the geometries sampled, allowing calculation of a barrier of 2 kcal/mol for the open-to-closed transition in the absence of ligands. MD simulations of fully liganded MshA revealed a smaller sampling of the dihedral angles. The localized protein fluctuation changes suggest that UDP-GlcNAc binding activates the motions of loops in the 1-l- myo -inositol-1-phosphate (I1P)-binding site despite little change in the interactions with UDP-GlcNAc. Circular dichroism, intrinsic fluorescence spectroscopy, and mutagenesis studies were used to confirm the ligand-induced structural changes in MshA. The results support a proposed mechanism where UDP-GlcNAc binds with rigid interactions to the C-terminal domain of MshA and activates flexible loops in the N-terminal domain for binding and positioning of I1P. This model can be used for future structure-based drug development of inhibitors of the mycothiol biosynthetic pathway. |
| المشرفين على المادة: | 0 (mycothiol) EC 2.4.- (Glycosyltransferases) 0 (Ligands) 0 (Inositol Phosphates) 58-98-0 (Uridine Diphosphate) K848JZ4886 (Cysteine) 0 (Glycopeptides) 4L6452S749 (Inositol) |
| تواريخ الأحداث: | Date Created: 20240320 Date Completed: 20240403 Latest Revision: 20240403 |
| رمز التحديث: | 20240403 |
| DOI: | 10.1021/acs.biochem.3c00561 |
| PMID: | 38507812 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-4995 |
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| DOI: | 10.1021/acs.biochem.3c00561 |