Structure of an acetylating aldehyde dehydrogenase from the thermophilic ethanologen Geobacillus thermoglucosidasius
Acetylating aldehyde dehydrogenases (AcAldDH) catalyse the acetylation of Coenzyme‐A (CoA), or in reverse generate acetaldehyde from Acetyl‐CoA using NADH as a co‐factor. This article reports the expression, purification, enzyme assay, and X‐ray crystal structures of an AcAldDH from Geobacillus ther...
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Published in | Protein science Vol. 25; no. 11; pp. 2045 - 2053 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
United States
Wiley Subscription Services, Inc
01.11.2016
John Wiley and Sons Inc |
Subjects | |
Online Access | Get full text |
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Summary: | Acetylating aldehyde dehydrogenases (AcAldDH) catalyse the acetylation of Coenzyme‐A (CoA), or in reverse generate acetaldehyde from Acetyl‐CoA using NADH as a co‐factor. This article reports the expression, purification, enzyme assay, and X‐ray crystal structures of an AcAldDH from Geobacillus thermoglucosidasius (GtAcAldDH) to 2.1Å and in complex with CoA and NAD+ to 4.0Å. In the structure, the AcAldDH forms a close‐knit dimer, similar to that seen in other Alcohol Dehydrogenase (ADH) structures. In GtAcAldDH, these dimers associate via their N‐termini to form weakly interacting tetramers. This mode of tetrameric association is also seen in an unpublished AcAldDH deposited in the PDB, but is in contrast to all other ADH structures, (including the one other published AcAldDH found in a bacterial microcompartment), in which the dimers bury a large surface area including the C‐termini. This novel mode of association sequesters the active sites and potentially reactive acyl‐enzyme intermediates in the center of the tetramer. In other respects, the structure is very similar to the other AcAldDH, binding the cofactors in a corresponding fashion. This similarity enabled the identification of a shortened substrate cavity in G. thermoglucosidasius AcAldDH, explaining the limitations on the length of substrate accepted by the enzyme.
PDB Code(s): 5J78; 5J71 |
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Bibliography: | Grant sponsor: TMO Renewables Ltd and the Biotechnology and Biological Sciences Research Council (UK) (a CASE studentship for JE). Conflict of interests: All authors declare that they have no competing interests in relation to the publication of this paper, that no funding source will gain or lose financially through publication of this paper, and that they have no personal financial interests that would be affected by this publication. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0961-8368 1469-896X |
DOI: | 10.1002/pro.3027 |