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

Job F, Settele F, Lorey S, Rundfeldt C, Baumann L, Beck-Sickinger AG, Haupts U, Lilie H, Bosse-Doenecke E - FEBS Open Bio (2015)

Bottom Line: 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.Furthermore, intravenous application to mice at high concentrations did not induce any detectable effect on cytokine levels or hematological parameters.

View Article: PubMed Central - PubMed

Affiliation: Institute for Biochemistry and Biotechnology/Technical Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Straße 3, D-06120 Halle (Saale), 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.

No MeSH data available.


Related in: MedlinePlus

Blood and serum accumulation of ubiquitin. (A) 125I-SIB-labeled ubiquitin (R&D Systems) or (B) F45W di-ubiquitin (Scil Proteins) was injected into the tail vein of 7 weeks old, female CD1 mice. Blood samples of indicated bleeding time points were collected from three non-anaesthetized mice. After analysis of whole blood and serum in a gamma counter, radioactivity was calculated and quantified as equivalent of protein provided as ng/total blood or serum. Error bars represent SD.
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f0010: Blood and serum accumulation of ubiquitin. (A) 125I-SIB-labeled ubiquitin (R&D Systems) or (B) F45W di-ubiquitin (Scil Proteins) was injected into the tail vein of 7 weeks old, female CD1 mice. Blood samples of indicated bleeding time points were collected from three non-anaesthetized mice. After analysis of whole blood and serum in a gamma counter, radioactivity was calculated and quantified as equivalent of protein provided as ng/total blood or serum. Error bars represent SD.

Mentions: Comparison of radioactivity measured in whole blood and the corresponding serum samples revealed accumulation of both proteins in the serum (Fig. 2A and B), i.e. there is no measurable cellular uptake of ubiquitin from the serum.


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

Job F, Settele F, Lorey S, Rundfeldt C, Baumann L, Beck-Sickinger AG, Haupts U, Lilie H, Bosse-Doenecke E - FEBS Open Bio (2015)

Blood and serum accumulation of ubiquitin. (A) 125I-SIB-labeled ubiquitin (R&D Systems) or (B) F45W di-ubiquitin (Scil Proteins) was injected into the tail vein of 7 weeks old, female CD1 mice. Blood samples of indicated bleeding time points were collected from three non-anaesthetized mice. After analysis of whole blood and serum in a gamma counter, radioactivity was calculated and quantified as equivalent of protein provided as ng/total blood or serum. Error bars represent SD.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4522466&req=5

f0010: Blood and serum accumulation of ubiquitin. (A) 125I-SIB-labeled ubiquitin (R&D Systems) or (B) F45W di-ubiquitin (Scil Proteins) was injected into the tail vein of 7 weeks old, female CD1 mice. Blood samples of indicated bleeding time points were collected from three non-anaesthetized mice. After analysis of whole blood and serum in a gamma counter, radioactivity was calculated and quantified as equivalent of protein provided as ng/total blood or serum. Error bars represent SD.
Mentions: Comparison of radioactivity measured in whole blood and the corresponding serum samples revealed accumulation of both proteins in the serum (Fig. 2A and B), i.e. there is no measurable cellular uptake of ubiquitin from the serum.

Bottom Line: 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.Furthermore, intravenous application to mice at high concentrations did not induce any detectable effect on cytokine levels or hematological parameters.

View Article: PubMed Central - PubMed

Affiliation: Institute for Biochemistry and Biotechnology/Technical Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Straße 3, D-06120 Halle (Saale), 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.

No MeSH data available.


Related in: MedlinePlus