2cfa: Difference between revisions
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==Overview== | ==Overview== | ||
By using biochemical and structural analyses, we have investigated the, catalytic mechanism of the recently discovered flavin-dependent, thymidylate synthase ThyX from Paramecium bursaria chlorella virus-1, (PBCV-1). Site-directed mutagenesis experiments have identified several, residues implicated in either NADPH oxidation or deprotonation activity of, PBCV-1 ThyX. Chemical modification by diethyl pyrocarbonate and mass, spectroscopic analyses identified a histidine residue (His53) crucial for, NADPH oxidation and located in the vicinity of the redox active N-5 atom, of the FAD ring system. Moreover, we observed that the conformation of, active site key residues of PBCV-1 ThyX differs from earlier reported ThyX, structures, suggesting structural changes during catalysis. Steady-state, . | By using biochemical and structural analyses, we have investigated the, catalytic mechanism of the recently discovered flavin-dependent, thymidylate synthase ThyX from Paramecium bursaria chlorella virus-1, (PBCV-1). Site-directed mutagenesis experiments have identified several, residues implicated in either NADPH oxidation or deprotonation activity of, PBCV-1 ThyX. Chemical modification by diethyl pyrocarbonate and mass, spectroscopic analyses identified a histidine residue (His53) crucial for, NADPH oxidation and located in the vicinity of the redox active N-5 atom, of the FAD ring system. Moreover, we observed that the conformation of, active site key residues of PBCV-1 ThyX differs from earlier reported ThyX, structures, suggesting structural changes during catalysis. Steady-state, kinetic analyses support a reaction mechanism where ThyX catalysis, proceeds via formation of distinct ternary complexes without formation of, a methyl enzyme intermediate. | ||
==About this Structure== | ==About this Structure== | ||
2CFA is a | 2CFA is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Paramecium_bursaria_chlorella_virus_1 Paramecium bursaria chlorella virus 1] with CME and FAD as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Thymidylate_synthase_(FAD) Thymidylate synthase (FAD)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.1.1.148 2.1.1.148] Structure known Active Site: AC1. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2CFA OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: tscp]] | [[Category: tscp]] | ||
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Revision as of 10:33, 5 November 2007
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STRUCTURE OF VIRAL FLAVIN-DEPENDANT THYMIDYLATE SYNTHASE THYX
Overview
By using biochemical and structural analyses, we have investigated the, catalytic mechanism of the recently discovered flavin-dependent, thymidylate synthase ThyX from Paramecium bursaria chlorella virus-1, (PBCV-1). Site-directed mutagenesis experiments have identified several, residues implicated in either NADPH oxidation or deprotonation activity of, PBCV-1 ThyX. Chemical modification by diethyl pyrocarbonate and mass, spectroscopic analyses identified a histidine residue (His53) crucial for, NADPH oxidation and located in the vicinity of the redox active N-5 atom, of the FAD ring system. Moreover, we observed that the conformation of, active site key residues of PBCV-1 ThyX differs from earlier reported ThyX, structures, suggesting structural changes during catalysis. Steady-state, kinetic analyses support a reaction mechanism where ThyX catalysis, proceeds via formation of distinct ternary complexes without formation of, a methyl enzyme intermediate.
About this Structure
2CFA is a Protein complex structure of sequences from Paramecium bursaria chlorella virus 1 with CME and FAD as ligands. Active as Thymidylate synthase (FAD), with EC number 2.1.1.148 Structure known Active Site: AC1. Full crystallographic information is available from OCA.
Reference
Catalytic mechanism and structure of viral flavin-dependent thymidylate synthase ThyX., Graziani S, Bernauer J, Skouloubris S, Graille M, Zhou CZ, Marchand C, Decottignies P, van Tilbeurgh H, Myllykallio H, Liebl U, J Biol Chem. 2006 Aug 18;281(33):24048-57. Epub 2006 May 17. PMID:16707489
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Proteopedia Page Contributors and Editors (what is this?)
- Pages with broken file links
- Paramecium bursaria chlorella virus 1
- Protein complex
- Thymidylate synthase (FAD)
- Bernauer, J.
- Decottignies, P.
- Graille, M.
- Graziani, S.
- Liebl, U.
- Marchand, C.
- Myllykallio, H.
- Skouloubris, S.
- Tilbeurgh, H.Van.
- Zhou, C.Z.
- CME
- FAD
- Fdts
- Flavin dependent thymidylate synthase fad
- Flavoprotein
- Methyltransferase
- Nucleotide biosynthesis
- Paramecium bursaria chlorella virus-1
- Thyx
- Transferase
- Tscp
