skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Purification, crystallization and preliminary X-ray characterization of the human GTP fucose pyrophosphorylase

Abstract

The human GTP fucose pyrophosphohydrolase protein has been crystallized via the hanging-drop technique over a reservoir of polyethylene glycol (MW 8000) and ethylene glycol. The orthorhombic crystals diffract to 2.8 Å resolution. The human nucleotide-sugar metabolizing enzyme GTP fucose pyrophosphorylase (GFPP) has been purified to homogeneity by an affinity chromatographic procedure that utilizes a novel nucleoside analog. This new purification regime results in a protein preparation that produces significantly better crystals than traditional purification methods. The purified 66.6 kDa monomeric protein has been crystallized via hanging-drop vapor diffusion at 293 K. Crystals of the native enzyme diffract to 2.8 Å and belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}. There is a single GFPP monomer in the asymmetric unit, giving a Matthews coefficient of 2.38 Å{sup 3} Da{sup −1} and a solvent content of 48.2%. A complete native data set has been collected as a first step in determining the three-dimensional structure of this enzyme.

Authors:
;  [1]
  1. Department of Chemistry and Biochemistry, University of Maryland, Baltimore County, 1000 Hilltop Circle, Chemistry 405C, Baltimore, MD 21250 (United States)
Publication Date:
OSTI Identifier:
22356329
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 4; Other Information: PMCID: PMC2222559; PMID: 16582493; PUBLISHER-ID: bw5131; OAI: oai:pubmedcentral.nih.gov:2222559; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; AFFINITY; CRYSTALLIZATION; CRYSTALS; DIFFUSION; MONOMERS; RESOLUTION; SACCHAROSE; SOLVENTS; SPACE GROUPS; VAPORS

Citation Formats

Quirk, Stephen, and Seley-Radtke, Katherine L., E-mail: kseley@umbc.edu. Purification, crystallization and preliminary X-ray characterization of the human GTP fucose pyrophosphorylase. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106008529.
Quirk, Stephen, & Seley-Radtke, Katherine L., E-mail: kseley@umbc.edu. Purification, crystallization and preliminary X-ray characterization of the human GTP fucose pyrophosphorylase. United Kingdom. doi:10.1107/S1744309106008529.
Quirk, Stephen, and Seley-Radtke, Katherine L., E-mail: kseley@umbc.edu. Sat . "Purification, crystallization and preliminary X-ray characterization of the human GTP fucose pyrophosphorylase". United Kingdom. doi:10.1107/S1744309106008529.
@article{osti_22356329,
title = {Purification, crystallization and preliminary X-ray characterization of the human GTP fucose pyrophosphorylase},
author = {Quirk, Stephen and Seley-Radtke, Katherine L., E-mail: kseley@umbc.edu},
abstractNote = {The human GTP fucose pyrophosphohydrolase protein has been crystallized via the hanging-drop technique over a reservoir of polyethylene glycol (MW 8000) and ethylene glycol. The orthorhombic crystals diffract to 2.8 Å resolution. The human nucleotide-sugar metabolizing enzyme GTP fucose pyrophosphorylase (GFPP) has been purified to homogeneity by an affinity chromatographic procedure that utilizes a novel nucleoside analog. This new purification regime results in a protein preparation that produces significantly better crystals than traditional purification methods. The purified 66.6 kDa monomeric protein has been crystallized via hanging-drop vapor diffusion at 293 K. Crystals of the native enzyme diffract to 2.8 Å and belong to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}. There is a single GFPP monomer in the asymmetric unit, giving a Matthews coefficient of 2.38 Å{sup 3} Da{sup −1} and a solvent content of 48.2%. A complete native data set has been collected as a first step in determining the three-dimensional structure of this enzyme.},
doi = {10.1107/S1744309106008529},
journal = {Acta Crystallographica. Section F},
number = Pt 4,
volume = 62,
place = {United Kingdom},
year = {Sat Apr 01 00:00:00 EST 2006},
month = {Sat Apr 01 00:00:00 EST 2006}
}