Mapping the Interaction of B Cell Leukemia 3 (BCL-3) and Nuclear Factor κB (NF-κB) p50 Identifies a BCL-3-mimetic Anti-inflammatory Peptide.
The NF-κB transcriptional response is tightly regulated by a number of processes including the phosphorylation, ubiquitination, and subsequent proteasomal degradation of NF-κB subunits. The IκB family protein BCL-3 stabilizes a NF-κB p50 homodimer·DNA complex through inhibition of p50 ubiquitination. This complex inhibits the binding of the transcriptionally active NF-κB subunits p65 and c-Rel on the promoters of NF-κB target genes and functions to suppress inflammatory gene expression. We have previously shown that the direct interaction between p50 and BCL-3 is required for BCL-3-mediated inhibition of pro-inflammatory gene expression. In this study we have used immobilized peptide array technology to define regions of BCl-3 that mediate interaction with p50 homodimers. Our data show that BCL-3 makes extensive contacts with p50 homodimers and in particular with ankyrin repeats (ANK) 1, 6, and 7, and the N-terminal region of Bcl-3. Using these data we have designed a BCL-3 mimetic peptide based on a region of the ANK1 of BCL-3 that interacts with p50 and shares low sequence similarity with other IκB proteins. When fused to a cargo carrying peptide sequence this BCL-3-derived peptide, but not a mutated peptide, inhibited Toll-like receptor-induced cytokine expression in vitro. The BCL-3 mimetic peptide was also effective in preventing inflammation in vivo in the carrageenan-induced paw edema mouse model. This study demonstrates that therapeutic strategies aimed at mimicking the functional activity of BCL-3 may be effective in the treatment of inflammatory disease.
Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, College of Medicine, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom.
Patricia E
PE
Collins
Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, College of Medicine, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom.
Gianluca
G
Grassia
Department of Biochemistry, University College Cork, Cork, Ireland.
Amy
A
Colleran
Department of Life Sciences, and Materials and Surface Science Institute, University of Limerick, Limerick, Ireland.
Patrick A
PA
Kiely
Department of Pharmacy, University of Napoli Federico II, Naples 80131, Italy.
Armando
A
Ialenti
Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, College of Medicine, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom; Department of Pharmacy, University of Napoli Federico II, Naples 80131, Italy.
Pasquale
P
Maffia
Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, College of Medicine, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom. Electronic address: ruaidhri.carmody@glasgow.ac.uk.
Ruaidhrí J
RJ
Carmody
D000818:Animals; D017089:Ankyrin Repeat; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology; D000081242:B-Cell Lymphoma 3 Protein; D040761:Biomimetic Materials / Q000737:chemistry / Q000494:pharmacology; D004195:Disease Models, Animal; D004487:Edema / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D052002:NF-kappa B p50 Subunit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D000081242:B-Cell Lymphoma 3 Protein; C000631344:BCL3 protein, human; C000631345:Bcl3 protein, mouse; D052002:NF-kappa B p50 Subunit; C489548:NFKB1 protein, human; D010455:Peptides; D011518:Proto-Oncogene Proteins; D014157:Transcription Factors; C073701:Nfkb1 protein, mouse
BCL-3; NF-κB; inflammation; peptide array; peptides; transcription