Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase.
N(2)-(2-Carboxyethyl)arginine synthase (CEAS), an unusual thiamin diphosphate (ThDP)-dependent enzyme, catalyses the committed step in the biosynthesis of the b-lactamase inhibitor clavulanic acid in Streptomyces clavuligerus. Crystal structures of tetrameric CEAS-ThDP in complex with the substrate...
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| Format: | Journal article |
| Language: | English |
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2009
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| _version_ | 1797102394464534528 |
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| author | Caines, M Sorensen, J Schofield, C |
| author_facet | Caines, M Sorensen, J Schofield, C |
| author_sort | Caines, M |
| collection | OXFORD |
| description | N(2)-(2-Carboxyethyl)arginine synthase (CEAS), an unusual thiamin diphosphate (ThDP)-dependent enzyme, catalyses the committed step in the biosynthesis of the b-lactamase inhibitor clavulanic acid in Streptomyces clavuligerus. Crystal structures of tetrameric CEAS-ThDP in complex with the substrate analogues 5-guanidinovaleric acid (GVA) and tartrate, and a structure reflecting a possible enol(ate)-ThDP reaction intermediate are described. The structures suggest overlapping binding sites for the substrates D-glyceraldehyde-3-phosphate (D-G3P) and L-arginine, and are consistent with the proposed CEAS mechanism in which D-G3P binds at the active site and reacts to form an alpha,beta-unsaturated intermediate,which subsequently undergoes (1,4)-Michael addition with the alpha-amino group of L-arginine. Additional solution studies are presented which probe the amino acid substrate tolerance of CEAS, providing further insight into the L-arginine binding site. These findings may facilitate the engineering of CEAS towards the synthesis of alternative beta-amino acid products. |
| first_indexed | 2024-03-07T06:05:24Z |
| format | Journal article |
| id | oxford-uuid:eda9a713-81df-42c0-accc-ce0c5c396c7a |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:05:24Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | oxford-uuid:eda9a713-81df-42c0-accc-ce0c5c396c7a2022-03-27T11:26:47ZStructural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:eda9a713-81df-42c0-accc-ce0c5c396c7aEnglishSymplectic Elements at Oxford2009Caines, MSorensen, JSchofield, CN(2)-(2-Carboxyethyl)arginine synthase (CEAS), an unusual thiamin diphosphate (ThDP)-dependent enzyme, catalyses the committed step in the biosynthesis of the b-lactamase inhibitor clavulanic acid in Streptomyces clavuligerus. Crystal structures of tetrameric CEAS-ThDP in complex with the substrate analogues 5-guanidinovaleric acid (GVA) and tartrate, and a structure reflecting a possible enol(ate)-ThDP reaction intermediate are described. The structures suggest overlapping binding sites for the substrates D-glyceraldehyde-3-phosphate (D-G3P) and L-arginine, and are consistent with the proposed CEAS mechanism in which D-G3P binds at the active site and reacts to form an alpha,beta-unsaturated intermediate,which subsequently undergoes (1,4)-Michael addition with the alpha-amino group of L-arginine. Additional solution studies are presented which probe the amino acid substrate tolerance of CEAS, providing further insight into the L-arginine binding site. These findings may facilitate the engineering of CEAS towards the synthesis of alternative beta-amino acid products. |
| spellingShingle | Caines, M Sorensen, J Schofield, C Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase. |
| title | Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase. |
| title_full | Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase. |
| title_fullStr | Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase. |
| title_full_unstemmed | Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase. |
| title_short | Structural and mechanistic studies on N(2)-(2-carboxyethyl)arginine synthase. |
| title_sort | structural and mechanistic studies on n 2 2 carboxyethyl arginine synthase |
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