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MBP‐binding DARPins facilitate the crystallization of an MBP fusion protein
Author(s) -
Gumpena Rajesh,
Lountos George T.,
Waugh David S.
Publication year - 2018
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x18009901
Subject(s) - maltose binding protein , fusion protein , ankyrin repeat , protein crystallization , crystallization , crystallography , fusion , escherichia coli , chemistry , biochemistry , biophysics , biology , computational biology , recombinant dna , linguistics , philosophy , organic chemistry , gene
The production of high‐quality crystals is the main bottleneck in determining the structures of proteins using X‐ray crystallography. In addition to being recognized as a very effective solubility‐enhancing fusion partner, Escherichia coli maltose‐binding protein (MBP) has also been successfully employed as a `fixed‐arm' crystallization chaperone in more than 100 cases. Here, it is reported that designed ankyrin‐repeat proteins (DARPins) that bind with high affinity to MBP can promote the crystallization of an MBP fusion protein when the fusion protein alone fails to produce diffraction‐quality crystals. As a proof of principle, three different co‐crystal structures of MBP fused to the catalytic domain of human dual‐specificity phosphatase 1 in complex with DARPins are reported.