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Title: 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 colimaltose-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:
; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1506492
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section F, Structural Biology Communications
Additional Journal Information:
Journal Volume: 74; Journal Issue: 9; Journal ID: ISSN 2053-230X
Publisher:
International Union of Crystallography
Country of Publication:
United States
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 States: N. p., 2018. Web. doi:10.1107/S2053230X18009901.
Gumpena, Rajesh, Lountos, George T., & Waugh, David S. MBP-binding DARPins facilitate the crystallization of an MBP fusion protein. United States. doi:10.1107/S2053230X18009901.
Gumpena, Rajesh, Lountos, George T., and Waugh, David S. Wed . "MBP-binding DARPins facilitate the crystallization of an MBP fusion protein". United States. doi:10.1107/S2053230X18009901.
@article{osti_1506492,
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 colimaltose-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},
issn = {2053-230X},
number = 9,
volume = 74,
place = {United States},
year = {2018},
month = {8}
}

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