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Updated: Jun 17, 2026

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
Published on: July 21, 2021
Identification and characterization of endonuclein binding proteins: evidence of modulatory effects on signal
Maja Ludvigsen1, Morten Østergaard, Henrik Vorum
1Department of Medical Biochemistry, Aarhus University, Ole Worms Allé 3, Building 1170, Aarhus, DK-8000 Aarhus C, Denmark. maja@biokemi.au.dk
Background:
We have previously identified endonuclein as a cell cycle regulated WD-repeat protein that is up-regulated in adenocarcinoma of the pancreas. Now, we aim to investigate its biomedical functions.
Results:
Using the cDNA encoding human endonuclein, we have expressed and purified the recombinant protein from Escherichia coli using metal affinity chromatography. The recombinant protein was immobilized to a column and by affinity chromatography several interacting proteins were purified from several litres of placenta tissue extract. After chromatography the eluted proteins were further separated by two-dimensional gel electrophoresis and identified by tandem mass spectrometry. The interacting proteins were identified as; Tax interaction protein 1 (TIP-1), Aalpha fibrinogen transcription factor (P16/SSBP1), immunoglobulin heavy chain binding protein (BiP), human ER-associated DNAJ (HEDJ/DNAJB11), endonuclein interaction protein 8 (EIP-8), and pregnancy specific beta-1 glycoproteins (PSGs). Surface plasmon resonance analysis and confocal fluorescence microscopy were used to further characterize the interactions.
Conclusions:
Our results demonstrate that endonuclein interacts with several proteins indicating a broad function including signal transduction and chaperone activity.
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