1mp5
Y177F VARIANT OF S. ENTERICA RmlA
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OverviewOverview
In vitro "glycorandomization" is a chemoenzymatic approach for generating, diverse libraries of glycosylated biomolecules based on natural product, scaffolds. This technology makes use of engineered variants of specific, enzymes affecting metabolite glycosylation, particularly, nucleotidylyltransferases and glycosyltransferases. To expand the, repertoire of UDP/dTDP sugars readily available for glycorandomization, we, now report a structure-based engineering approach to increase the, diversity of alpha-d-hexopyranosyl phosphates accepted by Salmonella, enterica LT2 alpha-d-glucopyranosyl phosphate thymidylyltransferase, (E(p)). This article highlights the design rationale, determined substrate, specificity, and structural elucidation of three "designed" mutations, illustrating both the success and unexpected outcomes from this type of, approach. In addition, a single amino acid substitution in the, substrate-binding pocket (L89T) was found to significantly increase the, set of alpha-d-hexopyranosyl phosphates accepted by E(p) to include, alpha-d-allo-, alpha-d-altro-, and alpha-d-talopyranosyl phosphate. In, aggregate, our results provide valuable blueprints for altering, nucleotidylyltransferase specificity by design, which is the first step, toward in vitro glycorandomization.
About this StructureAbout this Structure
1MP5 is a Single protein structure of sequence from Salmonella enterica with UPG as ligand. Active as Glucose-1-phosphate thymidylyltransferase, with EC number 2.7.7.24 Full crystallographic information is available from OCA.
ReferenceReference
Expanding pyrimidine diphosphosugar libraries via structure-based nucleotidylyltransferase engineering., Barton WA, Biggins JB, Jiang J, Thorson JS, Nikolov DB, Proc Natl Acad Sci U S A. 2002 Oct 15;99(21):13397-402. Epub 2002 Oct 8. PMID:12374866
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