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.
- Authors:
- Publication Date:
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1467532
- Resource Type:
- Published Article
- Journal Name:
- Acta Crystallographica Section F Structural Biology Communications
- Additional Journal Information:
- Journal Name: Acta Crystallographica Section F Structural Biology Communications Journal Volume: 74 Journal Issue: 9; 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.
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.
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.
@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}
}
DOI: 10.1107/S2053230X18009901