Chapter title |
The "Sneaking-Ligand" Approach: Cell-Type Specific Inhibition of the Classical NF-κB Pathway.
|
---|---|
Chapter number | 33 |
Book title |
NF-kappa B
|
Published in |
Methods in molecular biology, February 2015
|
DOI | 10.1007/978-1-4939-2422-6_33 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2421-9, 978-1-4939-2422-6
|
Authors |
Bettina Sehnert, Harald Burkhardt, Stefan Dübel, Reinhard E Voll, Reinhard E. Voll, Sehnert, Bettina, Burkhardt, Harald, Dübel, Stefan, Voll, Reinhard E. |
Editors |
Michael J. May |
Abstract |
The intracellular delivery of molecules across the plasma membrane represents a major obstacle. The conjugation of cell-permeable peptides (CPPs) to proteins promotes the uptake and internalization. However, uptake of CPPs is receptor independent and not cell-type specific. Recently, we established the "sneaking-ligand" approach which is based on multimodular recombinant fusion proteins that consist of three modules connected with serine-glycine linkers. Module one is responsible for receptor-mediated endocytosis; module two supports translocation into the cytoplasm so that the effector module three can interact with its binding partner in the cytoplasm. For NF-κB inhibition, we described an NF-κB inhibitor that targets selectively the activated endothelium via an oligopeptide motif. Upon E-selectin-mediated endocytosis, the Pseudomonas exotoxin A domain II (ETAII) translocates the NEMO-binding peptide to the cytoplasm interfering with IκB kinase complex assembly. Inflammatory autoimmune diseases are triggered, but also resolved by a variety of cell types. Therefore, the inhibition of NF-κB should be restricted to those cells that are crucially involved in the pathogenesis of inflammatory diseases. A general blockade of NF-κB may result in severe immunosuppression and possibly in organ dysfunction or damage. The "sneaking-ligand" approach could minimize the risks of therapeutic interventions and identify disease-relevant cell types. Here we describe the recombinant expression and purification of the E-selectin-specific "sneaking-ligand construct" (SLC1) and its ability to inhibit cytokine-induced NF-κB activation in vitro. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 5 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 2 | 40% |
Researcher | 1 | 20% |
Student > Doctoral Student | 1 | 20% |
Unknown | 1 | 20% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 1 | 20% |
Agricultural and Biological Sciences | 1 | 20% |
Immunology and Microbiology | 1 | 20% |
Chemistry | 1 | 20% |
Unknown | 1 | 20% |