Abstract
The methodology of protein crystallography provides a number of potential bottlenecks. Here, an approach to successful structure solution of a difficult heterodimeric complex of two human proteins, paraspeckle component 1 (PSPC1) and non-POU domain-containing octamer-binding protein (NONO), that are involved in gene regulation and the structural integrity of nuclear bodies termed paraspeckles is described. With the aid of bioinformatic predictions and systematic screening of a panel of constructs, bottlenecks of protein solubility, crystallization, crystal quality and crystallographic pseudosymmetry were overcome in order to produce crystals that ultimately revealed the structure.
© 2011 International Union of Crystallography. Printed in Singapore – all rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Computational Biology
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Crystallization
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Crystallography, X-Ray* / methods
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DNA-Binding Proteins
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Gene Expression Regulation
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Humans
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Intranuclear Inclusion Bodies / chemistry*
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Intranuclear Inclusion Bodies / genetics
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Intranuclear Inclusion Bodies / metabolism
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Multiprotein Complexes / chemistry*
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Multiprotein Complexes / metabolism
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Nuclear Matrix-Associated Proteins / chemistry*
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Nuclear Matrix-Associated Proteins / metabolism
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Nuclear Proteins / chemistry*
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Nuclear Proteins / metabolism
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Octamer Transcription Factors / chemistry*
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Octamer Transcription Factors / metabolism
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Protein Multimerization
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RNA-Binding Proteins / chemistry*
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RNA-Binding Proteins / metabolism
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Solubility
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Stereoisomerism
Substances
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DNA-Binding Proteins
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Multiprotein Complexes
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NONO protein, human
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Nuclear Matrix-Associated Proteins
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Nuclear Proteins
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Octamer Transcription Factors
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PSPC1 protein, human
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RNA-Binding Proteins