Source:http://linkedlifedata.com/resource/pubmed/id/15542052
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
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| pubmed:dateCreated |
2004-11-15
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| pubmed:databankReference | |
| pubmed:abstractText |
The naturally occurring, volatile sesquiterpene hydrocarbon germacrene D has strong effects on insect behaviour and genes encoding enzymes that produce this compound are of interest in the study of plant-insect interactions and in a number of biotechnological approaches to pest control. Goldenrod, Solidago canadensis, is unusual in that it produces both enantiomers of germacrene D. Two new sesquiterpene synthase cDNAs, designated Sc11 and Sc19, have been isolated from goldenrod and functional expression in Escherichia coli identified Sc11 as (+)-germacrene D synthase and Sc19 as (-)-germacrene D synthase. Thus, the enantiomers of germacrene D are the products of separate, but closely related (85% amino-acid identity), enzymes. Unlike other sesquiterpene synthases and the related monoterpene synthases and prenyl transferases, which contain the characteristic amino-acid motif DDXX(D,E), Sc11 is unusual in that this motif occurs as (303)NDTYD. Mutagenesis of this motif to (303)DDTYD gave rise to an enzyme that fully retained (+)-germacrene D synthase activity. The converse mutation in Sc19 (D303N) resulted in a less efficient but functional enzyme. Mutagenesis of position 303 to glutamate in both enzymes resulted in loss of activity. These results indicate that the magnesium ion-binding role of the first aspartate in the DDXXD motif may not be as critical as previously thought. Further amino-acid sequence comparisons and molecular modelling of the enzyme structures revealed that very subtle changes to the active site of this family of enzymes are required to alter the reaction pathway to form, in this case, different enantiomers from the same enzyme-bound carbocationic intermediate.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Alkyl and Aryl Transferases,
http://linkedlifedata.com/resource/pubmed/chemical/Aspartic Acid,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Terpenes,
http://linkedlifedata.com/resource/pubmed/chemical/germacrene D synthase
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| pubmed:status |
MEDLINE
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| pubmed:month |
Dec
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| pubmed:issn |
0003-9861
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
15
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| pubmed:volume |
432
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
136-44
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:15542052-Alkyl and Aryl Transferases,
pubmed-meshheading:15542052-Amino Acid Motifs,
pubmed-meshheading:15542052-Amino Acid Sequence,
pubmed-meshheading:15542052-Aspartic Acid,
pubmed-meshheading:15542052-Binding Sites,
pubmed-meshheading:15542052-Cells, Cultured,
pubmed-meshheading:15542052-Cloning, Molecular,
pubmed-meshheading:15542052-Computer Simulation,
pubmed-meshheading:15542052-Enzyme Activation,
pubmed-meshheading:15542052-Models, Chemical,
pubmed-meshheading:15542052-Models, Molecular,
pubmed-meshheading:15542052-Molecular Sequence Data,
pubmed-meshheading:15542052-Protein Binding,
pubmed-meshheading:15542052-Protein Conformation,
pubmed-meshheading:15542052-Protein Engineering,
pubmed-meshheading:15542052-Recombinant Proteins,
pubmed-meshheading:15542052-Solidago,
pubmed-meshheading:15542052-Structure-Activity Relationship,
pubmed-meshheading:15542052-Substrate Specificity,
pubmed-meshheading:15542052-Terpenes
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| pubmed:year |
2004
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| pubmed:articleTitle |
Enantiospecific (+)- and (-)-germacrene D synthases, cloned from goldenrod, reveal a functionally active variant of the universal isoprenoid-biosynthesis aspartate-rich motif.
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| pubmed:affiliation |
Rothamsted Research, Harpenden, Herts, AL5 2JQ, United Kingdom.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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