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PDBsum entry 4yyc
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References listed in PDB file
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Key reference
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Title
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Protein purification and crystallization artifacts: the tale usually not told.
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Authors
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E.Niedzialkowska,
O.Gasiorowska,
K.B.Handing,
K.A.Majorek,
P.J.Porebski,
I.G.Shabalin,
E.Zasadzinska,
M.Cymborowski,
W.Minor.
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Ref.
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Protein Sci, 2016,
25,
720-733.
[DOI no: ]
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PubMed id
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Abstract
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The misidentification of a protein sample, or contamination of a sample with the
wrong protein, may be a potential reason for the non-reproducibility of
experiments. This problem may occur in the process of heterologous
overexpression and purification of recombinant proteins, as well as purification
of proteins from natural sources. If the contaminated or misidentified sample is
used for crystallization, in many cases the problem may not be detected until
structures are determined. In the case of functional studies, the problem may
not be detected for years. Here several procedures that can be successfully used
for the identification of crystallized protein contaminants, including: (i) a
lattice parameter search against known structures, (ii) sequence or fold
identification from partially built models, and (iii) molecular replacement with
common contaminants as search templates have been presented. A list of common
contaminant structures to be used as alternative search models was provided.
These methods were used to identify four cases of purification and
crystallization artifacts. This report provides troubleshooting pointers for
researchers facing difficulties in phasing or model building.
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