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Title: Purification, crystallization and preliminary X-ray diffraction studies to near-atomic resolution of dihydrodipicolinate synthase from methicillin-resistant Staphylococcus aureus

Abstract

Dihydrodipicolinate synthase (DHDPS), an enzyme of the lysine-biosynthetic pathway, is a promising target for antibiotic development against pathogenic bacteria. Here, the expression, purification, crystallization and preliminary diffraction analysis to 1.45 Å resolution of DHDPS from methicillin-resistant S. aureus is reported. In recent years, dihydrodipicolinate synthase (DHDPS; EC 4.2.1.52) has received considerable attention from both mechanistic and structural viewpoints. DHDPS is part of the diaminopimelate pathway leading to lysine, coupling (S)-aspartate-β-semialdehyde with pyruvate via a Schiff base to a conserved active-site lysine. In this paper, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DHDPS from methicillin-resistant Staphylococcus aureus, an important bacterial pathogen, are reported. The enzyme was crystallized in a number of forms, predominantly from PEG precipitants, with the best crystal diffracting to beyond 1.45 Å resolution. The space group was P1 and the unit-cell parameters were a = 65.4, b = 67.6, c = 78.0 Å, α = 90.1, β = 68.9, γ = 72.3°. The crystal volume per protein weight (V{sub M}) was 2.34 Å{sup 3} Da{sup −1}, with an estimated solvent content of 47% for four monomers per asymmetric unit. The structure of the enzyme will help to guide the design of novel therapeutics againstmore » the methicillin-resistant S. aureus pathogen.« less

Authors:
; ;  [1];  [2];  [1]
  1. Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, Victoria 3010 (Australia)
  2. Centre for Structural Biology, Institute of Fundamental Sciences, Massey University, Palmerston North (New Zealand)
Publication Date:
OSTI Identifier:
22360598
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section F
Additional Journal Information:
Journal Volume: 64; Journal Issue: Pt 7; Other Information: PMCID: PMC2443978; PMID: 18607102; PUBLISHER-ID: nj5016; OAI: oai:pubmedcentral.nih.gov:2443978; Copyright (c) International Union of Crystallography 2008; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1744-3091
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; ANTIBIOTICS; COUPLING; CRYSTALLIZATION; CRYSTALS; DESIGN; LYSINE; MONOMERS; RESOLUTION; SCHIFF BASES; SOLVENTS; SPACE GROUPS; STAPHYLOCOCCUS; WEIGHT; X-RAY DIFFRACTION

Citation Formats

Burgess, Benjamin R., Dobson, Renwick C. J.,, Dogovski, Con, Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010, Jameson, Geoffrey B., Parker, Michael W., Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010, St Vincents Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065, Perugini, Matthew A., E-mail: rdobson@unimelb.edu.au, and Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010. Purification, crystallization and preliminary X-ray diffraction studies to near-atomic resolution of dihydrodipicolinate synthase from methicillin-resistant Staphylococcus aureus. United Kingdom: N. p., 2008. Web. doi:10.1107/S1744309108016746.
Burgess, Benjamin R., Dobson, Renwick C. J.,, Dogovski, Con, Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010, Jameson, Geoffrey B., Parker, Michael W., Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010, St Vincents Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065, Perugini, Matthew A., E-mail: rdobson@unimelb.edu.au, & Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010. Purification, crystallization and preliminary X-ray diffraction studies to near-atomic resolution of dihydrodipicolinate synthase from methicillin-resistant Staphylococcus aureus. United Kingdom. https://doi.org/10.1107/S1744309108016746
Burgess, Benjamin R., Dobson, Renwick C. J.,, Dogovski, Con, Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010, Jameson, Geoffrey B., Parker, Michael W., Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010, St Vincents Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065, Perugini, Matthew A., E-mail: rdobson@unimelb.edu.au, and Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010. 2008. "Purification, crystallization and preliminary X-ray diffraction studies to near-atomic resolution of dihydrodipicolinate synthase from methicillin-resistant Staphylococcus aureus". United Kingdom. https://doi.org/10.1107/S1744309108016746.
@article{osti_22360598,
title = {Purification, crystallization and preliminary X-ray diffraction studies to near-atomic resolution of dihydrodipicolinate synthase from methicillin-resistant Staphylococcus aureus},
author = {Burgess, Benjamin R. and Dobson, Renwick C. J., and Dogovski, Con and Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010 and Jameson, Geoffrey B. and Parker, Michael W. and Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010 and St Vincents Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065 and Perugini, Matthew A., E-mail: rdobson@unimelb.edu.au and Bio21 Molecular Science and Biotechnology Institute, 30 Flemington Road, Parkville, Victoria 3010},
abstractNote = {Dihydrodipicolinate synthase (DHDPS), an enzyme of the lysine-biosynthetic pathway, is a promising target for antibiotic development against pathogenic bacteria. Here, the expression, purification, crystallization and preliminary diffraction analysis to 1.45 Å resolution of DHDPS from methicillin-resistant S. aureus is reported. In recent years, dihydrodipicolinate synthase (DHDPS; EC 4.2.1.52) has received considerable attention from both mechanistic and structural viewpoints. DHDPS is part of the diaminopimelate pathway leading to lysine, coupling (S)-aspartate-β-semialdehyde with pyruvate via a Schiff base to a conserved active-site lysine. In this paper, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DHDPS from methicillin-resistant Staphylococcus aureus, an important bacterial pathogen, are reported. The enzyme was crystallized in a number of forms, predominantly from PEG precipitants, with the best crystal diffracting to beyond 1.45 Å resolution. The space group was P1 and the unit-cell parameters were a = 65.4, b = 67.6, c = 78.0 Å, α = 90.1, β = 68.9, γ = 72.3°. The crystal volume per protein weight (V{sub M}) was 2.34 Å{sup 3} Da{sup −1}, with an estimated solvent content of 47% for four monomers per asymmetric unit. The structure of the enzyme will help to guide the design of novel therapeutics against the methicillin-resistant S. aureus pathogen.},
doi = {10.1107/S1744309108016746},
url = {https://www.osti.gov/biblio/22360598}, journal = {Acta Crystallographica. Section F},
issn = {1744-3091},
number = Pt 7,
volume = 64,
place = {United Kingdom},
year = {Tue Jul 01 00:00:00 EDT 2008},
month = {Tue Jul 01 00:00:00 EDT 2008}
}