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Title: Expression, purification, crystallization and preliminary X-ray diffraction of a novel Nitrosomonas europaea cytochrome, cytochrome P460

Abstract

Cytochrome P460 from N. europaea, a novel mono-heme protein containing an unusual lysine cross-link to the porphyrin ring, has been recombinantly expressed and purified from E. coli and crystallized. The crystals belong to the trigonal space group P3{sub 1/2}21, with unit-cell parameters a = b = 53.3, c = 127.1 Å, one monomer in the asymmetric unit and diffract to 1.7 Å on a Cu Kα rotating-anode X-ray source. Cytochrome P460 from Nitrosomonas europaea, a novel mono-heme protein containing an unusual cross-link between a conserved lysine and the porphyrin ring, has been recombinantly expressed and purified from Escherichia coli. The protein crystallizes readily and diffraction to 1.7 Å has been obtained in-house. The crystals belong to the trigonal space group P3{sub 1/2}21, with unit-cell parameters a = b = 53.3, c = 127.1 Å, and contain one monomer in the asymmetric unit.

Authors:
; ; ;  [1]
  1. Department of Biochemistry, Molecular Biology and Biophysics, The University of Minnesota, Minneapolis, Minnesota (United States)
Publication Date:
OSTI Identifier:
22356341
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 4; Other Information: PMCID: PMC2222571; PMID: 16582494; PUBLISHER-ID: gx5094; OAI: oai:pubmedcentral.nih.gov:2222571; 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; ANODES; CRYSTALLIZATION; CRYSTALS; ESCHERICHIA COLI; HEME; LYSINE; MONOMERS; PROTEINS; RINGS; SPACE GROUPS; X-RAY DIFFRACTION; X-RAY SOURCES

Citation Formats

Elmore, Bradley O., Pearson, Arwen R., Wilmot, Carrie M., and Hooper, Alan B., E-mail: hooper@cbs.umn.edu. Expression, purification, crystallization and preliminary X-ray diffraction of a novel Nitrosomonas europaea cytochrome, cytochrome P460. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106008785.
Elmore, Bradley O., Pearson, Arwen R., Wilmot, Carrie M., & Hooper, Alan B., E-mail: hooper@cbs.umn.edu. Expression, purification, crystallization and preliminary X-ray diffraction of a novel Nitrosomonas europaea cytochrome, cytochrome P460. United Kingdom. doi:10.1107/S1744309106008785.
Elmore, Bradley O., Pearson, Arwen R., Wilmot, Carrie M., and Hooper, Alan B., E-mail: hooper@cbs.umn.edu. Sat . "Expression, purification, crystallization and preliminary X-ray diffraction of a novel Nitrosomonas europaea cytochrome, cytochrome P460". United Kingdom. doi:10.1107/S1744309106008785.
@article{osti_22356341,
title = {Expression, purification, crystallization and preliminary X-ray diffraction of a novel Nitrosomonas europaea cytochrome, cytochrome P460},
author = {Elmore, Bradley O. and Pearson, Arwen R. and Wilmot, Carrie M. and Hooper, Alan B., E-mail: hooper@cbs.umn.edu},
abstractNote = {Cytochrome P460 from N. europaea, a novel mono-heme protein containing an unusual lysine cross-link to the porphyrin ring, has been recombinantly expressed and purified from E. coli and crystallized. The crystals belong to the trigonal space group P3{sub 1/2}21, with unit-cell parameters a = b = 53.3, c = 127.1 Å, one monomer in the asymmetric unit and diffract to 1.7 Å on a Cu Kα rotating-anode X-ray source. Cytochrome P460 from Nitrosomonas europaea, a novel mono-heme protein containing an unusual cross-link between a conserved lysine and the porphyrin ring, has been recombinantly expressed and purified from Escherichia coli. The protein crystallizes readily and diffraction to 1.7 Å has been obtained in-house. The crystals belong to the trigonal space group P3{sub 1/2}21, with unit-cell parameters a = b = 53.3, c = 127.1 Å, and contain one monomer in the asymmetric unit.},
doi = {10.1107/S1744309106008785},
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}
}
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  • No abstract prepared.
  • Sec2p is an essential yeast gene and is part of the cell polarization process that leads to budding. The Nterminal domain of sec2p (Sec2pN) the guanine-nucleotide exchange factor for sec4p has been expressed in Escherichia coli, purified, and crystallized. Crystals belong to the space group P21 with unit cell dimensions 178.1 x 98.4 x 180.0 {angstrom}, {beta} = 91.7{sup o}, and diffract synchrotron-generated X-rays to better than 3.6 {angstrom} resolution. Pseudo-precession plots reveal a Laue symmetry of 2/m, corresponding to the aforementioned space group, and unusual weak diffraction in the -5--7 {angstrom} resolution range. The Matthews number calculations for amore » typical crystal density suggest a range of 28 to 64 molecules per asymmetric unit. Self-rotation and native Patterson calculations demonstrate a pure helical array of protein subunits. Based on the X-ray diffraction data analysis and amino-acid sequence alignments, the paper presents a hypothetical model of the exchange domain of sec2p as a pair of coiled-coil helices that binds to sec4p and facilitates nucleotide disassociation.« less