The soluble Y115E–Y117E variant of human glutaminyl cyclase is a valid target for X‐ray and NMR screening of inhibitors against Alzheimer disease. Issue 8 (1st August 2015)

• Record Type: Journal Article
• Authors: DiPisa, Flavio; Pozzi, Cecilia; Benvenuti, Manuela; Andreini, Matteo; Marconi, Guido; Mangani, Stefano
• Is Part Of: Acta crystallographica. Volume 71:Issue 8(2015:Aug.)
• Page Start: 986
• Page End: 992
• Publication Date: 2015-08-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X15010389
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Recent developments in molecular pathology and genetics have allowed the identification of human glutaminyl cyclase (hQC) among the abnormal proteins involved in many neurodegenerative disorders. Difficulties in obtaining large quantities of pure protein may limit the use of crystallographic screening for drug development on this target. Site‐directed mutagenesis experiments have led to the identification of some solvent‐exposed residues that are absolutely critical to achieve increased solubility and to avoid precipitation of the enzyme in inclusion bodies when expressed in <italic>Escherichia coli</italic>. The designed variant Y115E–Y117E has been found to be able to provide large amounts of monodisperse, pure hQC from an <italic>E. coli</italic> expression system. To validate the use of the artificial construct as a target for large‐scale X‐ray and NMR screening campaigns in the search for new inhibitors of hQC, the X‐ray crystal structures of the hQC Y115E–Y117E variant and of its adduct with the inhibitor PBD‐150 were determined.</p> </abstract>