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Title: Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6–8

Abstract

The crystallization of human complement regulator FH-678{sub 402H} with a glycosaminoglycan analogue is described. Human plasma protein complement factor H (FH) is an inhibitor of the spontaneously activated alternative complement pathway. An allotypic variant of FH, 402His, has been associated with age-related macular degeneration, the leading cause of blindness in the elderly. Crystals of FH domains 6–8 (FH678) containing 402His have been grown in the presence of a polyanionic sucrose octasulfate ligand (an analogue of the natural glycosaminoglycan ligands of FH) using both native and selenomethionine-derivatized protein. Native data sets diffracting to 2.3 Å and SeMet data sets of up to 2.8 Å resolution have been collected. An anomalous difference Patterson map reveals self- and cross-peaks from two incorporated Se atoms. The corresponding selenium substructure has been solved.

Authors:
; ;  [1];  [2];  [3];  [4];  [2];  [1]
  1. The Sir William Dunn School of Pathology, The University of Oxford, South Parks Road, Oxford OX1 3RE (United Kingdom)
  2. Faculty of Life Sciences, Manchester University, Michael Smith Building, Oxford Road, Manchester M13 9PT (United Kingdom)
  3. Medical Biomics Centre, St George’s, University of London, Cranmer Terrace, London SW17 0RE (United Kingdom)
  4. The MRC Immunochemistry Unit, The University of Oxford, South Parks Road, Oxford OX1 3RE (United Kingdom)
Publication Date:
OSTI Identifier:
22360345
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 63; Journal Issue: Pt 6; Other Information: PMCID: PMC2335081; PMID: 17554167; PUBLISHER-ID: ll5115; OAI: oai:pubmedcentral.nih.gov:2335081; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; ATOMS; CRYSTALLIZATION; CRYSTALS; LIGANDS; PEAKS; PLASMA; RESOLUTION; SACCHAROSE; SELENIUM

Citation Formats

Prosser, Beverly E., Johnson, Steven, Roversi, Pietro, Clark, Simon J., Tarelli, Edward, Sim, Robert B., Day, Antony J., and Lea, Susan M., E-mail: susan.lea@bnc.ox.ac.uk. Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6–8. United Kingdom: N. p., 2007. Web. doi:10.1107/S1744309107020052.
Prosser, Beverly E., Johnson, Steven, Roversi, Pietro, Clark, Simon J., Tarelli, Edward, Sim, Robert B., Day, Antony J., & Lea, Susan M., E-mail: susan.lea@bnc.ox.ac.uk. Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6–8. United Kingdom. doi:10.1107/S1744309107020052.
Prosser, Beverly E., Johnson, Steven, Roversi, Pietro, Clark, Simon J., Tarelli, Edward, Sim, Robert B., Day, Antony J., and Lea, Susan M., E-mail: susan.lea@bnc.ox.ac.uk. Fri . "Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6–8". United Kingdom. doi:10.1107/S1744309107020052.
@article{osti_22360345,
title = {Expression, purification, cocrystallization and preliminary crystallographic analysis of sucrose octasulfate/human complement regulator factor H SCRs 6–8},
author = {Prosser, Beverly E. and Johnson, Steven and Roversi, Pietro and Clark, Simon J. and Tarelli, Edward and Sim, Robert B. and Day, Antony J. and Lea, Susan M., E-mail: susan.lea@bnc.ox.ac.uk},
abstractNote = {The crystallization of human complement regulator FH-678{sub 402H} with a glycosaminoglycan analogue is described. Human plasma protein complement factor H (FH) is an inhibitor of the spontaneously activated alternative complement pathway. An allotypic variant of FH, 402His, has been associated with age-related macular degeneration, the leading cause of blindness in the elderly. Crystals of FH domains 6–8 (FH678) containing 402His have been grown in the presence of a polyanionic sucrose octasulfate ligand (an analogue of the natural glycosaminoglycan ligands of FH) using both native and selenomethionine-derivatized protein. Native data sets diffracting to 2.3 Å and SeMet data sets of up to 2.8 Å resolution have been collected. An anomalous difference Patterson map reveals self- and cross-peaks from two incorporated Se atoms. The corresponding selenium substructure has been solved.},
doi = {10.1107/S1744309107020052},
journal = {Acta Crystallographica. Section F},
number = Pt 6,
volume = 63,
place = {United Kingdom},
year = {Fri Jun 01 00:00:00 EDT 2007},
month = {Fri Jun 01 00:00:00 EDT 2007}
}
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