1mp5

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{{STRUCTURE_1mp5| PDB=1mp5 | SCENE= }}
{{STRUCTURE_1mp5| PDB=1mp5 | SCENE= }}
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'''Y177F VARIANT OF S. ENTERICA RmlA'''
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===Y177F VARIANT OF S. ENTERICA RmlA===
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==Overview==
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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.
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(as it appears on PubMed at http://www.pubmed.gov), where 12374866 is the PubMed ID number.
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{{ABSTRACT_PUBMED_12374866}}
==About this Structure==
==About this Structure==
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[[Category: Thorson, J S.]]
[[Category: Thorson, J S.]]
[[Category: Transferase]]
[[Category: Transferase]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 01:32:56 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Jul 3 00:39:02 2008''

Revision as of 21:39, 2 July 2008

Template:STRUCTURE 1mp5

Y177F VARIANT OF S. ENTERICA RmlA

Template:ABSTRACT PUBMED 12374866

About this Structure

1MP5 is a Single protein structure of sequence from Salmonella enterica. Full crystallographic information is available from OCA.

Reference

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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