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Title: Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis

Abstract

Phosphopantetheine adenylyltransferase from En. faecalis was crystallized and X-ray diffraction data were collected to 2.70 Å resolution. Phosphopantetheine adenylyltransferase, an essential enzyme in the coenzyme A biosynthetic pathway, catalyzes the reversible transfer of an adenylyl group from ATP to 4′-phosphopantetheine, yielding 3′-dephospho-CoA and pyrophosphate. Enterococcus faecalis PPAT has been overexpressed in Escherichia coli as a fusion with a C-terminal purification tag and crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium HEPES pH 7.5, 0.8 M sodium dihydrogen phosphate and 0.8 M potassium dihydrogen phosphate. X-ray diffraction data were collected to 2.70 Å at 100 K. The crystals belong to the primitive tetragonal space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = b = 160.81, c = 225.68 Å. Four copies of the hexameric molecule are likely to be present in the asymmetric unit, giving a crystal volume per protein weight (V{sub M}) of 3.08 Å{sup 3} Da{sup −1} and a solvent content of 60.1%.

Authors:
; ; ;
Publication Date:
OSTI Identifier:
22356386
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section F
Additional Journal Information:
Journal Volume: 62; Journal Issue: Pt 11; Other Information: PMCID: PMC2225208; PMID: 17077496; PUBLISHER-ID: en5200; OAI: oai:pubmedcentral.nih.gov:2225208; Copyright (c) International Union of Crystallography 2006; 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; CRYSTALS; ESCHERICHIA COLI; MATHEMATICAL SOLUTIONS; MOLECULES; RESOLUTION; SODIUM; SOLUTIONS; SOLVENTS; SPACE GROUPS; WEIGHT; X-RAY DIFFRACTION

Citation Formats

Kang, Ji Yong, Lee, Hyung Ho, Yoon, Hye Jin, Kim, Hyoun Sook, and Suh, Se Won, E-mail: sewonsuh@snu.ac.kr. Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106041108.
Kang, Ji Yong, Lee, Hyung Ho, Yoon, Hye Jin, Kim, Hyoun Sook, & Suh, Se Won, E-mail: sewonsuh@snu.ac.kr. Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis. United Kingdom. https://doi.org/10.1107/S1744309106041108
Kang, Ji Yong, Lee, Hyung Ho, Yoon, Hye Jin, Kim, Hyoun Sook, and Suh, Se Won, E-mail: sewonsuh@snu.ac.kr. 2006. "Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis". United Kingdom. https://doi.org/10.1107/S1744309106041108.
@article{osti_22356386,
title = {Overexpression, crystallization and preliminary X-ray crystallographic analysis of phosphopantetheine adenylyltransferase from Enterococcus faecalis},
author = {Kang, Ji Yong and Lee, Hyung Ho and Yoon, Hye Jin and Kim, Hyoun Sook and Suh, Se Won, E-mail: sewonsuh@snu.ac.kr},
abstractNote = {Phosphopantetheine adenylyltransferase from En. faecalis was crystallized and X-ray diffraction data were collected to 2.70 Å resolution. Phosphopantetheine adenylyltransferase, an essential enzyme in the coenzyme A biosynthetic pathway, catalyzes the reversible transfer of an adenylyl group from ATP to 4′-phosphopantetheine, yielding 3′-dephospho-CoA and pyrophosphate. Enterococcus faecalis PPAT has been overexpressed in Escherichia coli as a fusion with a C-terminal purification tag and crystallized at 297 K using a reservoir solution consisting of 0.1 M sodium HEPES pH 7.5, 0.8 M sodium dihydrogen phosphate and 0.8 M potassium dihydrogen phosphate. X-ray diffraction data were collected to 2.70 Å at 100 K. The crystals belong to the primitive tetragonal space group P4{sub 1} (or P4{sub 3}), with unit-cell parameters a = b = 160.81, c = 225.68 Å. Four copies of the hexameric molecule are likely to be present in the asymmetric unit, giving a crystal volume per protein weight (V{sub M}) of 3.08 Å{sup 3} Da{sup −1} and a solvent content of 60.1%.},
doi = {10.1107/S1744309106041108},
url = {https://www.osti.gov/biblio/22356386}, journal = {Acta Crystallographica. Section F},
issn = {1744-3091},
number = Pt 11,
volume = 62,
place = {United Kingdom},
year = {Wed Nov 01 00:00:00 EST 2006},
month = {Wed Nov 01 00:00:00 EST 2006}
}