Overexpression, purification, crystallization, and preliminary X-ray characterization of a methionine sulfoxide reductase AB from Helicobacter pylori

Kitaik Lee, Hyun Sook Kim, Won Kyu Lee, Ah Reum Han, Jun Soo Kim, Kwang Yeon Hwang

Research output: Contribution to journalArticle

Abstract

The main function of methionine sulfoxide reductases (Msr) in many organisms is to protect cells against oxidative stress caused by the catalyzed reduction of oxidized methionine to normal methionine. In a few micro-organisms, the existence of Msr as a fusion protein on a single polypeptide, MsrAB, was reported. However, Msr generally exists as separate enzymes MsrA and MsrB. Here, MsrAB from Helicobacter pylori (HpMsrAB) was overexpressed in Escherichia coli, purified, and crystallized to determine its structure. HpMsrAB X-ray diffraction data were collected to the resolution of 3.3 Å, and the crystals were found to belong to the tetragonal space group P41212, with the unit cell parameters a =100.91, b =100.91, and c =160.08 Å. The crystals corresponded to 5.38 Å3 Da-1 of Matthews coefficient and 77.2% solvent content from the molecular replacement suggest that there is a single molecule in an asymmetric unit. Due to their unusually high solvent content, diffraction of these crystals only reach a resolution of 3.3 Å. A preliminary solution was determined by molecular replacement. Further refinement of the structure is in progress.

Original languageEnglish
Pages (from-to)23-26
Number of pages4
JournalJournal of the Korean Society for Applied Biological Chemistry
Volume57
Issue number1
DOIs
Publication statusPublished - 2014 Feb 1

Fingerprint

Methionine Sulfoxide Reductases
Crystallization
Helicobacter pylori
Purification
X-Rays
X rays
Methionine
Crystals
Oxidative stress
X-Ray Diffraction
Escherichia coli
Oxidative Stress
Fusion reactions
Diffraction
X ray diffraction
Peptides
Molecules
Enzymes
methionine sulfoxide reductase
Proteins

Keywords

  • crystallization
  • fusion protein
  • pathogen
  • reactive oxygen species
  • reductase

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Organic Chemistry

Cite this

Overexpression, purification, crystallization, and preliminary X-ray characterization of a methionine sulfoxide reductase AB from Helicobacter pylori. / Lee, Kitaik; Kim, Hyun Sook; Lee, Won Kyu; Han, Ah Reum; Kim, Jun Soo; Hwang, Kwang Yeon.

In: Journal of the Korean Society for Applied Biological Chemistry, Vol. 57, No. 1, 01.02.2014, p. 23-26.

Research output: Contribution to journalArticle

@article{2cc94db8efb84c769e0517c4519d5870,
title = "Overexpression, purification, crystallization, and preliminary X-ray characterization of a methionine sulfoxide reductase AB from Helicobacter pylori",
abstract = "The main function of methionine sulfoxide reductases (Msr) in many organisms is to protect cells against oxidative stress caused by the catalyzed reduction of oxidized methionine to normal methionine. In a few micro-organisms, the existence of Msr as a fusion protein on a single polypeptide, MsrAB, was reported. However, Msr generally exists as separate enzymes MsrA and MsrB. Here, MsrAB from Helicobacter pylori (HpMsrAB) was overexpressed in Escherichia coli, purified, and crystallized to determine its structure. HpMsrAB X-ray diffraction data were collected to the resolution of 3.3 {\AA}, and the crystals were found to belong to the tetragonal space group P41212, with the unit cell parameters a =100.91, b =100.91, and c =160.08 {\AA}. The crystals corresponded to 5.38 {\AA}3 Da-1 of Matthews coefficient and 77.2{\%} solvent content from the molecular replacement suggest that there is a single molecule in an asymmetric unit. Due to their unusually high solvent content, diffraction of these crystals only reach a resolution of 3.3 {\AA}. A preliminary solution was determined by molecular replacement. Further refinement of the structure is in progress.",
keywords = "crystallization, fusion protein, pathogen, reactive oxygen species, reductase",
author = "Kitaik Lee and Kim, {Hyun Sook} and Lee, {Won Kyu} and Han, {Ah Reum} and Kim, {Jun Soo} and Hwang, {Kwang Yeon}",
year = "2014",
month = "2",
day = "1",
doi = "10.1007/s13765-013-4183-5",
language = "English",
volume = "57",
pages = "23--26",
journal = "Applied Biological Chemistry",
issn = "2468-0834",
publisher = "Springer Netherlands",
number = "1",

}

TY - JOUR

T1 - Overexpression, purification, crystallization, and preliminary X-ray characterization of a methionine sulfoxide reductase AB from Helicobacter pylori

AU - Lee, Kitaik

AU - Kim, Hyun Sook

AU - Lee, Won Kyu

AU - Han, Ah Reum

AU - Kim, Jun Soo

AU - Hwang, Kwang Yeon

PY - 2014/2/1

Y1 - 2014/2/1

N2 - The main function of methionine sulfoxide reductases (Msr) in many organisms is to protect cells against oxidative stress caused by the catalyzed reduction of oxidized methionine to normal methionine. In a few micro-organisms, the existence of Msr as a fusion protein on a single polypeptide, MsrAB, was reported. However, Msr generally exists as separate enzymes MsrA and MsrB. Here, MsrAB from Helicobacter pylori (HpMsrAB) was overexpressed in Escherichia coli, purified, and crystallized to determine its structure. HpMsrAB X-ray diffraction data were collected to the resolution of 3.3 Å, and the crystals were found to belong to the tetragonal space group P41212, with the unit cell parameters a =100.91, b =100.91, and c =160.08 Å. The crystals corresponded to 5.38 Å3 Da-1 of Matthews coefficient and 77.2% solvent content from the molecular replacement suggest that there is a single molecule in an asymmetric unit. Due to their unusually high solvent content, diffraction of these crystals only reach a resolution of 3.3 Å. A preliminary solution was determined by molecular replacement. Further refinement of the structure is in progress.

AB - The main function of methionine sulfoxide reductases (Msr) in many organisms is to protect cells against oxidative stress caused by the catalyzed reduction of oxidized methionine to normal methionine. In a few micro-organisms, the existence of Msr as a fusion protein on a single polypeptide, MsrAB, was reported. However, Msr generally exists as separate enzymes MsrA and MsrB. Here, MsrAB from Helicobacter pylori (HpMsrAB) was overexpressed in Escherichia coli, purified, and crystallized to determine its structure. HpMsrAB X-ray diffraction data were collected to the resolution of 3.3 Å, and the crystals were found to belong to the tetragonal space group P41212, with the unit cell parameters a =100.91, b =100.91, and c =160.08 Å. The crystals corresponded to 5.38 Å3 Da-1 of Matthews coefficient and 77.2% solvent content from the molecular replacement suggest that there is a single molecule in an asymmetric unit. Due to their unusually high solvent content, diffraction of these crystals only reach a resolution of 3.3 Å. A preliminary solution was determined by molecular replacement. Further refinement of the structure is in progress.

KW - crystallization

KW - fusion protein

KW - pathogen

KW - reactive oxygen species

KW - reductase

UR - http://www.scopus.com/inward/record.url?scp=84897748767&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84897748767&partnerID=8YFLogxK

U2 - 10.1007/s13765-013-4183-5

DO - 10.1007/s13765-013-4183-5

M3 - Article

AN - SCOPUS:84897748767

VL - 57

SP - 23

EP - 26

JO - Applied Biological Chemistry

JF - Applied Biological Chemistry

SN - 2468-0834

IS - 1

ER -