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MBP-binding DARPins facilitate the crystallization of an MBP fusion protein.

ABSTRACT: 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.

SUBMITTER: Gumpena R

PROVIDER: S-EPMC6130421 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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MBP-binding DARPins facilitate the crystallization of an MBP fusion protein.

Gumpena Rajesh R Lountos George T GT Waugh David S DS

Acta crystallographica. Section F, Structural biology communications 20180829 Pt 9

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 crysta ...[more]

PMID: 30198887

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