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Biochemical and biophysical comparison of the human and Drosophila melanogaster I kappa B kinase protein complexes suggest structural conservation
Author(s) -
Rogers W Eric,
Huxford Tom
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.603.22
Subject(s) - drosophila melanogaster , biology , schneider 2 cells , melanogaster , signal transduction , protein subunit , transcription factor , microbiology and biotechnology , gene , biochemistry , kinase , peptide sequence , iκb kinase , protein kinase a , peptide , genetics , rna interference , rna
The IMD signaling pathway in Drosophila melanogaster is responsible for the expression of antimicrobial peptide genes in response to detection and binding of products of Gram‐negative bacterial infection at the cell membrane. Several macromolecular components of the IMD signaling pathway have close structural and/or functional homologs within the canonical NF‐κB signal transduction pathway that regulates inflammation and cell survival in mammals. Chief among these is the IκB kinase complex (IKK), which functions in both humans and flies to activate transcription factor NF‐κB. In this study, we compare the biophysical and biochemical properties of the Drosophila melanogaster IKK complex with those of its human homolog. We find striking similarities in both solution properties and intra‐complex interactions. Size exclusion chromatography measurements indicate similar hydrodynamic radii for the enzymes. Multi‐angle light scattering reveals a similar shape independent molecular weight. Hydrogen/deuterium exchange‐mass spectrometry studies identify that the region that is most protected upon binding is at the C‐terminus of the human and Drosophila IKKbeta subunit. Interestingly, residues that map to theoretical ubiquitin‐like domain are also protected upon interaction with IKKgamma. These experiments suggest overall structural conservation between the human and Drosophila IKKbeta:IKKgamma complexes despite considerable amino acid sequence variation in the individual polypeptide subunits.