Exploring Calbindin-IMPase fusion proteins structure and activity

Noble, James W. and Atack, John R. ORCID: https://orcid.org/0000-0002-3410-791X 2022. Exploring Calbindin-IMPase fusion proteins structure and activity. Biochemistry and Biophysics Reports 30 , 101266. 10.1016/j.bbrep.2022.101266

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Abstract

Calbindin-D28k is a calcium binding protein that is highly expressed in the mammalian central nervous system. It has been reported that calbindin-D28k binds to and increases the activity of inositol Monophosphatase (IMPase). This is an enzyme that is involved in the homeostasis of the Inositol trisphosphate signalling cascade by catalysing the final dephosphorylation of inositol and has been implicated in the therapeutic mechanism of lithium treatment of bipolar disorder. Previously studies have shown that calbindin-D28k can increase IMPase activity by up to 250 hundred-fold. A preliminary in silico model was proposed for the interaction. Here, we aimed at exploring the shape and properties of the calbindin-IMPase complex to gain new insights on this biologically important interaction. We created several fusion constructs of calbindin-D28k and IMPase, connected by flexible amino acid linkers of different lengths and orientations to fuse the termini of the two proteins together. The resulting fusion proteins have activities 200%–400% higher the isolated wild-type IMPase. The constructs were characterized by small angle X-ray scattering to gain information on the overall shape of the complexes and validate the previous model. The fusion proteins form a V-shaped, elongated and less compact complex as compared to the model. Our results shed new light into this protein-protein interaction.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Additional Information:	This is an open access article under the CC-BY-NC-ND license.
ISSN:	2405-5808
Date of First Compliant Deposit:	9 June 2022
Date of Acceptance:	20 April 2022
Last Modified:	24 May 2023 00:20
URI:	https://orca.cardiff.ac.uk/id/eprint/150373

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