Ubiquitin is a versatile scaffold protein for the generation of molecules with de novo binding and advantageous drug-like properties

Florian Job; Florian Settele; Susan Lorey; Chris Rundfeldt; Lars Baumann; Annette G Beck-Sickinger; Ulrich Haupts; Hauke Lilie; Eva Bosse-Doenecke

doi:10.1016/j.fob.2015.07.002

Ubiquitin is a versatile scaffold protein for the generation of molecules with de novo binding and advantageous drug-like properties

FEBS Open Bio. 2015 Jul 10:5:579-93. doi: 10.1016/j.fob.2015.07.002. eCollection 2015.

Authors

Florian Job¹, Florian Settele², Susan Lorey², Chris Rundfeldt², Lars Baumann³, Annette G Beck-Sickinger³, Ulrich Haupts², Hauke Lilie¹, Eva Bosse-Doenecke²

Affiliations

¹ Institute for Biochemistry and Biotechnology/Technical Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Straße 3, D-06120 Halle (Saale), Germany.
² Scil Proteins GmbH, Heinrich-Damerow-Straße 1, D-06120 Halle (Saale), Germany.
³ Institute of Biochemistry, University of Leipzig, Brüderstraße 34, D-04103 Leipzig, Germany.

Abstract

In the search for effective therapeutic strategies, protein-based biologicals are under intense development. While monoclonal antibodies represent the majority of these drugs, other innovative approaches are exploring the use of scaffold proteins for the creation of binding molecules with tailor-made properties. Ubiquitin is especially suited for this strategy due to several key characteristics. Ubiquitin is a natural serum protein, 100% conserved across the mammalian class and possesses high thermal, structural and proteolytic stability. Because of its small size and lack of posttranslational modifications, it can be easily produced in Escherichia coli. In this work we provide evidence that ubiquitin is safe as tested experimentally in vivo. In contrast to previously published results, we show that, in our hands, ubiquitin does not act as a functional ligand of the chemokine receptor CXCR4. Cellular assays based on different signaling pathways of the receptor were conducted with the natural agonist SDF-1 as a benchmark. In none of the assays could a response to ubiquitin treatment be elicited. Furthermore, intravenous application to mice at high concentrations did not induce any detectable effect on cytokine levels or hematological parameters.

Keywords: 125I-SIB, N-succinimidyl 3-(125iodo)-benzoate; Biodistribution; CXCR4; CXCR4, CXC motif chemokine receptor 4; G-protein coupled receptor; ID, injected dose; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; MCV, mean cell volume; PBS, phosphate buffered saline; SDF-1; SDF-1, stromal cell-derived factor 1; Scaffold; Ubiquitin.