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

doi:10.1107/S2053230X18009901

Title: MBP-binding DARPins facilitate the crystallization of an MBP fusion protein

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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.

Authors:: Gumpena, Rajesh; Lountos, George T.; Waugh, David S.

Publication Date:: 2018-08-29

Sponsoring Org.:: USDOE

OSTI Identifier:: 1467532

Resource Type:: Published Article

Journal Name:: Acta Crystallographica Section F Structural Biology Communications

Additional Journal Information:: Journal Volume: 74; Journal Issue: 9; Related Information: CHORUS Timestamp: 2018-09-06 10:11:59; Journal ID: ISSN 2053-230X

Publisher:: International Union of Crystallography (IUCr)

Country of Publication:: United Kingdom

Language:: English

Citation Formats


                    Gumpena, Rajesh, Lountos, George T., and Waugh, David S. MBP-binding DARPins facilitate the crystallization of an MBP fusion protein.  United Kingdom: N. p., 2018. 
        Web.  doi:10.1107/S2053230X18009901.

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                    Gumpena, Rajesh, Lountos, George T., & Waugh, David S. MBP-binding DARPins facilitate the crystallization of an MBP fusion protein.  United Kingdom.  doi:10.1107/S2053230X18009901.

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                    Gumpena, Rajesh, Lountos, George T., and Waugh, David S. Wed .  
        "MBP-binding DARPins facilitate the crystallization of an MBP fusion protein".  United Kingdom.  doi:10.1107/S2053230X18009901.

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@article{osti_1467532,

  title        = {MBP-binding DARPins facilitate the crystallization of an MBP fusion protein},

  author       = {Gumpena, Rajesh and Lountos, George T. and Waugh, David S.},

  abstractNote = {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.},

  doi          = {10.1107/S2053230X18009901},

  journal      = {Acta Crystallographica Section F Structural Biology Communications},

  number       = 9,

  volume       = 74,

  place        = {United Kingdom},

  year         = {2018},

  month        = {8}

}

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DOI: 10.1107/S2053230X18009901

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