Cloning, purification and preliminary X-ray crystallographic analysis of a hypothetical protein, MJ0754, from Methanococcus jannaschii DSM 2661

Eun Hye Lee, Ki Hyun Nam, Kwang Yeon Hwang

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1 Citation (Scopus)

Abstract

The protein encoded by the MJ0754 gene from the archaeon Methanococcus jannaschii DSM 2661 is an unknown hypothetical protein. Two recombinant proteins, MJ0754 (residues 1-185) and MJ0754t (a truncated form of MJ0754, residues 11-185), were cloned from MJ0754, overexpressed as His-tag fusion proteins and purified. The crystals were found to grow under two different conditions and to have two different shapes. The crystal of MJ0754 belonged to space group P61, with unit-cell parameters a = b = 127.015, c = 48.929 Å, a calculated Matthews coefficient of 2.85 Å3 Da-1 and two molecules per asymmetric unit. The crystal of MJ0754t belonged to space group C2221, with unit-cell parameters a = 51.915, b = 79.122, c = 93.869 Å, a calculated Matthews coefficient of 2.41 Å3 Da -1 and one molecule per asymmetric unit. The SeMet-labelled P61 crystal diffracted to a resolution of 3.1 Å, while the native C2221 crystal diffracted to 1.3 Å resolution.

Original languageEnglish
Pages (from-to)1065-1067
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume65
Issue number10
DOIs
Publication statusPublished - 2009 Oct 29

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Methanocaldococcus
Cloning
purification
Purification
Organism Cloning
X-Rays
proteins
X rays
Crystals
crystals
Proteins
x rays
Archaea
Recombinant Proteins
Molecules
coefficients
cells
genes
molecules
Fusion reactions

Keywords

  • Ferritin-like diiron-carboxylate proteins
  • Hypothetical proteins
  • Methanococcus jannaschii
  • MJ0754

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Structural Biology
  • Genetics
  • Condensed Matter Physics

Cite this

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title = "Cloning, purification and preliminary X-ray crystallographic analysis of a hypothetical protein, MJ0754, from Methanococcus jannaschii DSM 2661",
abstract = "The protein encoded by the MJ0754 gene from the archaeon Methanococcus jannaschii DSM 2661 is an unknown hypothetical protein. Two recombinant proteins, MJ0754 (residues 1-185) and MJ0754t (a truncated form of MJ0754, residues 11-185), were cloned from MJ0754, overexpressed as His-tag fusion proteins and purified. The crystals were found to grow under two different conditions and to have two different shapes. The crystal of MJ0754 belonged to space group P61, with unit-cell parameters a = b = 127.015, c = 48.929 {\AA}, a calculated Matthews coefficient of 2.85 {\AA}3 Da-1 and two molecules per asymmetric unit. The crystal of MJ0754t belonged to space group C2221, with unit-cell parameters a = 51.915, b = 79.122, c = 93.869 {\AA}, a calculated Matthews coefficient of 2.41 {\AA}3 Da -1 and one molecule per asymmetric unit. The SeMet-labelled P61 crystal diffracted to a resolution of 3.1 {\AA}, while the native C2221 crystal diffracted to 1.3 {\AA} resolution.",
keywords = "Ferritin-like diiron-carboxylate proteins, Hypothetical proteins, Methanococcus jannaschii, MJ0754",
author = "Lee, {Eun Hye} and Nam, {Ki Hyun} and Hwang, {Kwang Yeon}",
year = "2009",
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T1 - Cloning, purification and preliminary X-ray crystallographic analysis of a hypothetical protein, MJ0754, from Methanococcus jannaschii DSM 2661

AU - Lee, Eun Hye

AU - Nam, Ki Hyun

AU - Hwang, Kwang Yeon

PY - 2009/10/29

Y1 - 2009/10/29

N2 - The protein encoded by the MJ0754 gene from the archaeon Methanococcus jannaschii DSM 2661 is an unknown hypothetical protein. Two recombinant proteins, MJ0754 (residues 1-185) and MJ0754t (a truncated form of MJ0754, residues 11-185), were cloned from MJ0754, overexpressed as His-tag fusion proteins and purified. The crystals were found to grow under two different conditions and to have two different shapes. The crystal of MJ0754 belonged to space group P61, with unit-cell parameters a = b = 127.015, c = 48.929 Å, a calculated Matthews coefficient of 2.85 Å3 Da-1 and two molecules per asymmetric unit. The crystal of MJ0754t belonged to space group C2221, with unit-cell parameters a = 51.915, b = 79.122, c = 93.869 Å, a calculated Matthews coefficient of 2.41 Å3 Da -1 and one molecule per asymmetric unit. The SeMet-labelled P61 crystal diffracted to a resolution of 3.1 Å, while the native C2221 crystal diffracted to 1.3 Å resolution.

AB - The protein encoded by the MJ0754 gene from the archaeon Methanococcus jannaschii DSM 2661 is an unknown hypothetical protein. Two recombinant proteins, MJ0754 (residues 1-185) and MJ0754t (a truncated form of MJ0754, residues 11-185), were cloned from MJ0754, overexpressed as His-tag fusion proteins and purified. The crystals were found to grow under two different conditions and to have two different shapes. The crystal of MJ0754 belonged to space group P61, with unit-cell parameters a = b = 127.015, c = 48.929 Å, a calculated Matthews coefficient of 2.85 Å3 Da-1 and two molecules per asymmetric unit. The crystal of MJ0754t belonged to space group C2221, with unit-cell parameters a = 51.915, b = 79.122, c = 93.869 Å, a calculated Matthews coefficient of 2.41 Å3 Da -1 and one molecule per asymmetric unit. The SeMet-labelled P61 crystal diffracted to a resolution of 3.1 Å, while the native C2221 crystal diffracted to 1.3 Å resolution.

KW - Ferritin-like diiron-carboxylate proteins

KW - Hypothetical proteins

KW - Methanococcus jannaschii

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