Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis
Cofactor F420 is an electron carrier with a major role in the oxidoreductive reactions of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB). A γ-glutamyl ligase catalyzes the final steps of the F420 biosynthesis pathway by successive additions of L-glutamate residues to F420...
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American Society for Biochemistry and Molecular Biology Inc.
2016
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| Online Access: | http://irep.iium.edu.my/50126/ http://irep.iium.edu.my/50126/7/50126-Elongation%20of%20the%20poly-%CE%B3-glutamate%20tail%20of.pdf http://irep.iium.edu.my/50126/4/50126-Elongation_of_the_poly_WOS.pdf |
| _version_ | 1848783581668704256 |
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| author | Bashiri, Ghader Mohamed Rehan, Aisyah Sreebhavan, Sreevalsan Baker, Heather M. Baker, Edward N. Squire, Christopher J. |
| author_facet | Bashiri, Ghader Mohamed Rehan, Aisyah Sreebhavan, Sreevalsan Baker, Heather M. Baker, Edward N. Squire, Christopher J. |
| author_sort | Bashiri, Ghader |
| building | IIUM Repository |
| collection | Online Access |
| description | Cofactor F420 is an electron carrier with a major role in the oxidoreductive reactions of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB). A γ-glutamyl ligase catalyzes the final steps of the F420 biosynthesis pathway by successive additions of L-glutamate residues to F420-0, producing a poly-γ-glutamate tail. The enzyme responsible for this reaction in Archaea (CofE) comprises a single domain and produces F420-2 as the major species. The homologous Mtb enzyme, FbiB, is a two-domain protein and produces F420 with predominantly 5-7 L-glutamate residues in the poly-γ-glutamate tail. The N-terminal domain of FbiB is homologous to CofE with an annotated γ-glutamyl ligase activity, whereas the C-terminal domain has sequence similarity to an FMN-dependent family of nitroreductase enzymes. Here we demonstrate that full-length FbiB adds multiple L-glutamate residues to F420-0 in vitro to produce F420-5 after 24 hours; communication between the two domains is critical for full γ-glutamyl ligase activity. We also present crystal structures of the C-terminal domain of FbiB in apo, F420-0 and FMN bound states, displaying distinct sites for F420-0 and FMN ligands that partially overlap. Finally, we discuss the features of a full-length structural model produced by small angle X-ray scattering (SAXS) and its implications for the role of N- and C-terminal domains in catalysis. |
| first_indexed | 2025-11-14T16:23:38Z |
| format | Article |
| id | iium-50126 |
| institution | International Islamic University Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-14T16:23:38Z |
| publishDate | 2016 |
| publisher | American Society for Biochemistry and Molecular Biology Inc. |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | iium-501262017-10-23T03:37:36Z http://irep.iium.edu.my/50126/ Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis Bashiri, Ghader Mohamed Rehan, Aisyah Sreebhavan, Sreevalsan Baker, Heather M. Baker, Edward N. Squire, Christopher J. Q Science (General) Cofactor F420 is an electron carrier with a major role in the oxidoreductive reactions of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB). A γ-glutamyl ligase catalyzes the final steps of the F420 biosynthesis pathway by successive additions of L-glutamate residues to F420-0, producing a poly-γ-glutamate tail. The enzyme responsible for this reaction in Archaea (CofE) comprises a single domain and produces F420-2 as the major species. The homologous Mtb enzyme, FbiB, is a two-domain protein and produces F420 with predominantly 5-7 L-glutamate residues in the poly-γ-glutamate tail. The N-terminal domain of FbiB is homologous to CofE with an annotated γ-glutamyl ligase activity, whereas the C-terminal domain has sequence similarity to an FMN-dependent family of nitroreductase enzymes. Here we demonstrate that full-length FbiB adds multiple L-glutamate residues to F420-0 in vitro to produce F420-5 after 24 hours; communication between the two domains is critical for full γ-glutamyl ligase activity. We also present crystal structures of the C-terminal domain of FbiB in apo, F420-0 and FMN bound states, displaying distinct sites for F420-0 and FMN ligands that partially overlap. Finally, we discuss the features of a full-length structural model produced by small angle X-ray scattering (SAXS) and its implications for the role of N- and C-terminal domains in catalysis. American Society for Biochemistry and Molecular Biology Inc. 2016-03-25 Article PeerReviewed application/pdf en http://irep.iium.edu.my/50126/7/50126-Elongation%20of%20the%20poly-%CE%B3-glutamate%20tail%20of.pdf application/pdf en http://irep.iium.edu.my/50126/4/50126-Elongation_of_the_poly_WOS.pdf Bashiri, Ghader and Mohamed Rehan, Aisyah and Sreebhavan, Sreevalsan and Baker, Heather M. and Baker, Edward N. and Squire, Christopher J. (2016) Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis. Journal of Biological Chemistry, 291 (13). pp. 6882-6894. ISSN 0021-9258 E-ISSN 1083-351X http://www.jbc.org/content/291/13/6882.abstract 10.1074/jbc.M115.689026 |
| spellingShingle | Q Science (General) Bashiri, Ghader Mohamed Rehan, Aisyah Sreebhavan, Sreevalsan Baker, Heather M. Baker, Edward N. Squire, Christopher J. Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis |
| title | Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis |
| title_full | Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis |
| title_fullStr | Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis |
| title_full_unstemmed | Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis |
| title_short | Elongation of the poly-γ-glutamate tail of F420 requires both domains of the F420:γ-glutamyl ligase (FbiB) of Mycobacterium tuberculosis |
| title_sort | elongation of the poly-γ-glutamate tail of f420 requires both domains of the f420:γ-glutamyl ligase (fbib) of mycobacterium tuberculosis |
| topic | Q Science (General) |
| url | http://irep.iium.edu.my/50126/ http://irep.iium.edu.my/50126/ http://irep.iium.edu.my/50126/ http://irep.iium.edu.my/50126/7/50126-Elongation%20of%20the%20poly-%CE%B3-glutamate%20tail%20of.pdf http://irep.iium.edu.my/50126/4/50126-Elongation_of_the_poly_WOS.pdf |