The refined crystal structure of Drosophila lebanonensis alcohol dehydrogenase at 1.9 å resolution 1 1Edited by R. Huber

Benach, J.; Atrian, Sı́lvia; Gonzàlez‐Duarte, Roser; Ladenstein, Rudolf

doi:10.1006/jmbi.1998.2015

Cited by 102 publications

(117 citation statements)

References 70 publications

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“…2A), consisting of a central ␤-sheet surrounded by ␣-helices and a typical nicotinamide coenzyme binding ␤␣␤␣␤ subdomain with a characteristic Gly-Xaa-Gly-Xaa-Xaa-Gly motif (position 13-18). Asp-37 confers specificity toward NAD binding, whereas the active site is characterized by a Ser-Tyr-Lys catalytic triad (12)(13)(14). These and other conserved SDR features are preserved in JGW, as shown in Fig.…”

Section: Resultsmentioning

confidence: 89%

“…2B) (12, 13). Two replacements, His-191 3 Gln (His191Gln) and Glu-205 3 Lys (Glu205Lys), lie between Thr-186 and Pro-210, a region that forms the flap guarding the entrance to the active site and that is known to be responsible for the different specificities among related enzymes (12,13). Two additional replacements, Ser215Pro and Leu120Met, contact residues within the Thr-186-Pro-210 region and also may influence specificity.…”

Section: Resultsmentioning

confidence: 99%

“…Molecular graphics were created with the program VMD. The template used to build the D. teissieri and D. yakuba three-dimensional model was the crystal structure of the orthologous enzyme D. melanogaster ADH (Protein Data Bank ID code 1MG5), and D. lebanonensis ADH (Protein Data Bank ID codes 1B14, 1B15, 1B16, and 1B2L) (12). When needed, data about the position of the coenzyme (NAD ϩ ) and the substrate were extracted from the crystal structures of binary and ternary enzyme complexes.…”

Section: Mapping Substitutions On the Three-dimensional Structures Ofmentioning

confidence: 99%

See 2 more Smart Citations

Evolving protein functional diversity in new genes of Drosophila

Zhang

Dean

Brunet

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

107

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The mechanism by which protein functional diversity expands is an important evolutionary issue. Studies of recently evolved chimeric genes permit direct investigation of the origin of new protein functions before they become obscured by subsequent evolution. Found in several African Drosophila species, jingwei (jgw), a recently evolved gene with a domain derived from the still extant short-chain alcohol dehydrogenase (ADH) through retroposition, provides an opportunity to examine this previously undescribed process directly. We expressed JGW proteins in a microbial expression system and, after purification, investigated their enzymatic properties. We found that, unexpectedly, positive Darwinian selection for amino acid replacements outside the active site of JGW produced a novel dehydrogenase with altered substrate specificity compared with the ancestral ADH. Instead of detoxifying and assimilating ethanol like its Adh parental gene, we observe that JGW efficiently utilizes long-chain primary alcohols found in hormone and pheromone metabolism. These data suggest that protein functional diversity can expand rapidly under the joint forces of exon shuffling, gene duplication, and natural selection.

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Section: Resultsmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Mapping Substitutions On the Three-dimensional Structures Ofmentioning

confidence: 99%

See 1 more Smart Citation

Evolving protein functional diversity in new genes of Drosophila

Zhang

Dean

Brunet

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

107

View full text Add to dashboard Cite

show abstract

“…The major proteins with a native alcohol-binding function are alcohol dehydrogenases, which invariably involve coordination of metal ions and other cofactors to the alcohol to perform an enzymatic reaction [14][15][16][17] . This mode of alcohol binding is unrelated to the non-enzymatic sites that occur in alcohol-sensitive ion-channels.…”

Section: Introductionmentioning

confidence: 99%

Structure of a specific alcohol-binding site defined by the odorant binding protein LUSH from Drosophila melanogaster

Kruse

Zhao

Smith

et al. 2003

Nat Struct Mol Biol

211

265

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SummaryWe have solved the high-resolution crystal structure of the Drosophila melanogaster alcoholbinding protein LUSH in the complexes it forms with a series of short chain n-alcohols. LUSH is the first known non-enzyme protein with a defined in vivo alcohol-binding function. The structure of LUSH reveals a set of molecular interactions that define a specific alcohol-binding site. A group of amino acids, Thr57, Ser52 and Thr48 form a network of concerted hydrogen bonds between the protein and the alcohol that provide a structural motif to increase alcohol binding affinity at this site. This motif appears to be conserved in a number of mammalian ligand-gated ion channels that are directly implicated in the pharmacological effects of alcohol. Further, these sequences are found in regions of ion-channels that are known to infer alcohol sensitivity. We suggest the alcohol-binding site in LUSH represents a general model for alcohol-binding sites in proteins.

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“…Although they are conserved between MtDH and SCR, their side chain conformations in the NADP-free crystal form differ dramatically from those in the MtDH-NADP + complex structure (PDB code: 1H5Q). The loop between bF and aG contains one small helix (aFG, residues 230-238), which is responsible for substrate specificity among the different enzymes within the SDR family (Ghosh et al 1994;Tanaka et al 1996;Benach et al 1998;Mazza et al 1998;Yamashita et al 1999). Furthermore, SCR contains an extended N-terminal peptide (i.e., residues 1-25) and a short helix (residues 26-30) projecting out from the core domain that might stabilize the oligomer (see below).…”

Section: Resultsmentioning

confidence: 99%

Crystal structure of a carbonyl reductase from Candida parapsilosis with anti‐Prelog stereospecificity

et al. 2008

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A novel short-chain (S)-1-phenyl-1,2-ethanediol dehydrogenase (SCR) from Candida parapsilosis exhibits coenzyme specificity for NADPH over NADH. It catalyzes an anti-Prelog type reaction to reduce 2-hydroxyacetophenone into (S)-1-phenyl-1,2-ethanediol. The coding gene was overexpressed in Escherichia coli and the purified protein was crystallized. The crystal structure of the apo-form was solved to 2.7 Å resolution. This protein forms a homo-tetramer with a broken 2-2-2 symmetry. The overall fold of each SCR subunit is similar to that of the known structures of other homologous alcohol dehydrogenases, although the latter usually form tetramers with perfect 2-2-2 symmetries. Additionally, in the apo-SCR structure, the entrance of the NADPH pocket is blocked by a surface loop. In order to understand the structure-function relationship of SCR, we carried out a number of mutagenesisenzymatic analyses based on the new structural information. First, mutations of the putative catalytic Ser-Tyr-Lys triad confirmed their functional role. Second, truncation of an N-terminal 31-residue peptide indicated its role in oligomerization, but not in catalytic activity. Similarly, a V270D point mutation rendered the SCR as a dimer, rather than a tetramer, without affecting the enzymatic activity. Moreover, the S67D/H68D double-point mutation inside the coenzyme-binding pocket resulted in a nearly 10-fold increase and a 20-fold decrease in the k cat /K M value when NADH and NADPH were used as cofactors, respectively, with k cat remaining essentially the same. This latter result provides a new example of a protein engineering approach to modify the coenzyme specificity in SCR and short-chain dehydrogenases/reductases in general.

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The refined crystal structure of Drosophila lebanonensis alcohol dehydrogenase at 1.9 å resolution 1 1Edited by R. Huber

Cited by 102 publications

References 70 publications

Evolving protein functional diversity in new genes of Drosophila

Evolving protein functional diversity in new genes of Drosophila

Structure of a specific alcohol-binding site defined by the odorant binding protein LUSH from Drosophila melanogaster

Crystal structure of a carbonyl reductase from Candida parapsilosis with anti‐Prelog stereospecificity

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