Mapping the conformational space accessible to BACE2 using surface mutants and cocrystals with Fab fragments, Fynomers and Xaperones

The aspartic protease BACE2 is responsible for the shedding of the transmembrane protein Tmem27 from the surface of pancreatic β‐cells, which leads to inactivation of the β‐cell proliferating activity of Tmem27. This role of BACE2 in the control of β‐cell maintenance suggests BACE2 as a drug target...

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Published in:	Acta crystallographica. Section D, Biological crystallography. Vol. 69; no. 6; pp. 1124 - 1137
Main Authors:	Banner, David W., Gsell, Bernard, Benz, Jörg, Bertschinger, Julian, Burger, Dominique, Brack, Simon, Cuppuleri, Simon, Debulpaep, Maja, Gast, Alain, Grabulovski, Dragan, Hennig, Michael, Hilpert, Hans, Huber, Walter, Kuglstatter, Andreas, Kusznir, Eric, Laeremans, Toon, Matile, Hugues, Miscenic, Christian, Rufer, Arne C., Schlatter, Daniel, Steyaert, Jan, Stihle, Martine, Thoma, Ralf, Weber, Martin, Ruf, Armin
Format:	Journal Article
Language:	English
Published:	5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01-06-2013 Wiley Subscription Services, Inc
Subjects:	Amyloid Precursor Protein Secretases - chemistry Amyloid Precursor Protein Secretases - genetics Amyloid Precursor Protein Secretases - metabolism Animals Antibodies Area Under Curve Aspartic Acid Endopeptidases - chemistry Aspartic Acid Endopeptidases - genetics Aspartic Acid Endopeptidases - metabolism Binders Catalytic Domain Crystallization Crystals drug discovery and design Drugs Enzymes Fragments Humans Immunoglobulin Fab Fragments - chemistry Immunoglobulin Fab Fragments - metabolism Insulin-Secreting Cells - enzymology Insulin-Secreting Cells - metabolism Mice Models, Molecular Mutagenesis Protein Conformation protein crystallization Proteins Surface chemistry Surface Plasmon Resonance X-Ray Diffraction protein crystallization drug discovery and design antibodies
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Summary:	The aspartic protease BACE2 is responsible for the shedding of the transmembrane protein Tmem27 from the surface of pancreatic β‐cells, which leads to inactivation of the β‐cell proliferating activity of Tmem27. This role of BACE2 in the control of β‐cell maintenance suggests BACE2 as a drug target for diabetes. Inhibition of BACE2 has recently been shown to lead to improved control of glucose homeostasis and to increased insulin levels in insulin‐resistant mice. BACE2 has 52% sequence identity to the well studied Alzheimer's disease target enzyme β‐secretase (BACE1). High‐resolution BACE2 structures would contribute significantly to the investigation of this enzyme as either a drug target or anti‐target. Surface mutagenesis, BACE2‐binding antibody Fab fragments, single‐domain camelid antibody VHH fragments (Xaperones) and Fyn‐kinase‐derived SH3 domains (Fynomers) were used as crystallization helpers to obtain the first high‐resolution structures of BACE2. Eight crystal structures in six different packing environments define an ensemble of low‐energy conformations available to the enzyme. Here, the different strategies used for raising and selecting BACE2 binders for cocrystallization are described and the crystallization success, crystal quality and the time and resources needed to obtain suitable crystals are compared.
Bibliography:	ArticleID:AYDTZ5028 ark:/67375/WNG-646ZCQHV-V istex:143FCA0148A2D7CBEF164F0D6B61022515A1F6E0 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	1399-0047 0907-4449 1399-0047
DOI:	10.1107/S0907444913006574