1mp4: Difference between revisions

Revision as of 01:32, 3 May 2008

W224H VARIANT OF S. ENTERICA RmlA

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

1MP4 is a Single protein structure of sequence from Salmonella enterica. 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 Page seeded by OCA on Sat May 3 01:32:46 2008

Proteopedia Page Contributors and Editors (what is this?)Proteopedia Page Contributors and Editors (what is this?)

OCA

@@ Line 1: / Line 1: @@
 [[Image:1mp4.jpg|left|200px]]
- {{Structure
+<!--
-|PDB= 1mp4 |SIZE=350|CAPTION= <scene name='initialview01'>1mp4</scene>, resolution 2.20&Aring;
+The line below this paragraph, containing "STRUCTURE_1mp4", creates the "Structure Box" on the page.
-|SITE=
+You may change the PDB parameter (which sets the PDB file loaded into the applet)
-|LIGAND= <scene name='pdbligand=UPG:URIDINE-5&#39;-DIPHOSPHATE-GLUCOSE'>UPG</scene>
+or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
-|ACTIVITY= <span class='plainlinks'>[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] </span>
+or leave the SCENE parameter empty for the default display.
-|GENE=
+-->
-|DOMAIN=
+{{STRUCTURE_1mp4|  PDB=1mp4  |  SCENE=  }}
-|RELATEDENTRY=[[1iim|1IIM]], [[1iin|1IIN]], [[1mp3|1MP3]], [[1mp5|1MP5]]
-|RESOURCES=<span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=1mp4 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=1mp4 OCA], [http://www.ebi.ac.uk/pdbsum/1mp4 PDBsum], [http://www.rcsb.org/pdb/explore.do?structureId=1mp4 RCSB]</span>
-}}
 '''W224H VARIANT OF S. ENTERICA RmlA'''
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 [[Category: Nikolov, D B.]]
 [[Category: Thorson, J S.]]
-[[Category: transferase]]
+[[Category: Transferase]]
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May  3 01:32:46 2008''
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Mar 30 22:19:08 2008''