1mp5: Difference between revisions
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New page: left|200px<br /><applet load="1mp5" size="450" color="white" frame="true" align="right" spinBox="true" caption="1mp5, resolution 2.75Å" /> '''Y177F VARIANT OF S. ... |
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[[Image:1mp5.jpg|left|200px]]<br /><applet load="1mp5" size=" | [[Image:1mp5.jpg|left|200px]]<br /><applet load="1mp5" size="350" color="white" frame="true" align="right" spinBox="true" | ||
caption="1mp5, resolution 2.75Å" /> | caption="1mp5, resolution 2.75Å" /> | ||
'''Y177F VARIANT OF S. ENTERICA RmlA'''<br /> | '''Y177F VARIANT OF S. ENTERICA RmlA'''<br /> | ||
==Overview== | ==Overview== | ||
In vitro "glycorandomization" is a chemoenzymatic approach for generating | 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 Structure== | ==About this Structure== | ||
1MP5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Salmonella_enterica Salmonella enterica] with UPG as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Glucose-1-phosphate_thymidylyltransferase Glucose-1-phosphate thymidylyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.24 2.7.7.24] Full crystallographic information is available from [http:// | 1MP5 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Salmonella_enterica Salmonella enterica] with <scene name='pdbligand=UPG:'>UPG</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Glucose-1-phosphate_thymidylyltransferase Glucose-1-phosphate thymidylyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.7.24 2.7.7.24] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1MP5 OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: Salmonella enterica]] | [[Category: Salmonella enterica]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Barton, W | [[Category: Barton, W A.]] | ||
[[Category: Biggins, J | [[Category: Biggins, J B.]] | ||
[[Category: Jiang, J.]] | [[Category: Jiang, J.]] | ||
[[Category: Nikolov, D | [[Category: Nikolov, D B.]] | ||
[[Category: Thorson, J | [[Category: Thorson, J S.]] | ||
[[Category: UPG]] | [[Category: UPG]] | ||
[[Category: transferase]] | [[Category: transferase]] | ||
''Page seeded by [http:// | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:57:35 2008'' | ||
