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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

IL-2 and IL-12 level after ubiquitin treatment. In a pilot acute toxicity study blood samples of rats (n = 8, 4 male and 4 female) treated with various doses of F45W di-ubiquitin (Scil Proteins, grey bars) or the vehicle control (white bars) were analyzed. (A and B) IL-2 and (C and D) IL-12 levels of male (A and C, n = 4) and female (B and D, n = 4) rats were determined with a bead array on a Cytomics FC 500 at indicated time points. Detection limit of the IL-2 assay was determined at 4.0 pg/ml. All measured values below this lower limit were set to 4.0 pg/ml. Error bars represent SD.
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f0035: IL-2 and IL-12 level after ubiquitin treatment. In a pilot acute toxicity study blood samples of rats (n = 8, 4 male and 4 female) treated with various doses of F45W di-ubiquitin (Scil Proteins, grey bars) or the vehicle control (white bars) were analyzed. (A and B) IL-2 and (C and D) IL-12 levels of male (A and C, n = 4) and female (B and D, n = 4) rats were determined with a bead array on a Cytomics FC 500 at indicated time points. Detection limit of the IL-2 assay was determined at 4.0 pg/ml. All measured values below this lower limit were set to 4.0 pg/ml. Error bars represent SD.


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)

IL-2 and IL-12 level after ubiquitin treatment. In a pilot acute toxicity study blood samples of rats (n = 8, 4 male and 4 female) treated with various doses of F45W di-ubiquitin (Scil Proteins, grey bars) or the vehicle control (white bars) were analyzed. (A and B) IL-2 and (C and D) IL-12 levels of male (A and C, n = 4) and female (B and D, n = 4) rats were determined with a bead array on a Cytomics FC 500 at indicated time points. Detection limit of the IL-2 assay was determined at 4.0 pg/ml. All measured values below this lower limit were set to 4.0 pg/ml. Error bars represent SD.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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

f0035: IL-2 and IL-12 level after ubiquitin treatment. In a pilot acute toxicity study blood samples of rats (n = 8, 4 male and 4 female) treated with various doses of F45W di-ubiquitin (Scil Proteins, grey bars) or the vehicle control (white bars) were analyzed. (A and B) IL-2 and (C and D) IL-12 levels of male (A and C, n = 4) and female (B and D, n = 4) rats were determined with a bead array on a Cytomics FC 500 at indicated time points. Detection limit of the IL-2 assay was determined at 4.0 pg/ml. All measured values below this lower limit were set to 4.0 pg/ml. Error bars represent SD.
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