Ivičak-Kocjan, K, Panter, G, Benčina, M*, Jerala, R*. Determination of the physiological 2:2 TLR5:flagellin activation stoichiometry revealed by the activity of a fusion receptor. (Biochem biophys res commun 2013, 435. [5222426], IF1.95). Project leader shares corresponding authorship. Authors determined the physiological stoichiometry of TLR5:flagellin activation by the use of a chimeric protein composed of an active flagellin fragment linked to the N-terminus of human TLR5 (SF-TLR5). This construct was constitutively active. Addition of wild-type hTLR5 substantially lowered autoactivation of SF-TLR5 in a concentration dependent manner, an effect which was reversible by the addition of exogenous S. typhimurium flagellin, indicating the biological activity of a TLR5:flagellin complex with a 2:2 stoichiometry. These results, in addition to the combinations of inactive P736H mutation within the BB-loop of the TIR domain of TLR5 and SF-TLR5, further confirm the mechanism of TLR5 activation. KIK, GB performed experiments, MB supervised work and analyzed data. MB and RJ wrote manuscript. The work was also presented as lecture: Molecular mechanism of TLR5 activation by flagellin (Meeting of the Slovenian Biochemical Society with International Participation [36980229]).
COBISS.SI-ID: 5222426
Pohar J, Pirher N, Benčina M, Manček Keber M, Jerala R*. The role of UNC93B1 protein in surface localization of TLR3 receptor and in cell priming to nucleic acid agonists. (J Biol Chem 2013, 288 [5143066] [IF4.651, 2 citations]) observed that poly(I:C) up-regulates transcription of UNC93B1 and promotes trafficking of TLR3 to the plasma membrane in human epithelial cell line. Further studies revealed that expression of UNC93B1 promotes trafficking of differentially glycosylated TLR3, but not other NAS TLRs, to the plasma membrane. Our findings identified TLR3 as the important regulator of UNC93B1 that in turn governs the responsiveness of all NAS TLRs.
COBISS.SI-ID: 5143066
Gradišar H, … Hafner Bratkovič I, … Jerala R*. Design of a single-chain polypeptide tetrahedron assembled from coiled-coil segments. (Nature Chem Biol 2013, 9 [5222682], [IF12.95, 8 citations] Authors present a strategy to design self-assembling polypeptide nanostructured polyhedra based on modularization using orthogonal dimerizing segments. They designed and experimentally demonstrated the formation of the tetrahedron that self-assembles from a single polypeptide chain comprising 12 concatenated coiled coil-forming segments separated by flexible peptide hinges. This design platform provides a foundation for constructing new topological polypeptide folds based on the set of orthogonal interacting polypeptide segments. The work was also presented as lectures: Imperial College, London, UK, 2013 [5340698], Alpbach, Austria 2013 [36977925], Málaga 2013 [5388826]
COBISS.SI-ID: 5222682