title: Structural characterization of the intra-membrane histidine kinase YbdK from Bacillus subtilis in DPC micelles.
abstract: Bacterial histidine kinases (HKs) play a critical role in signal transduction for cellular adaptation to environmental conditions and stresses. YbdK from Bacillus subtilis is a 320-residue intra-membrane sensing HK characterized by a short input domain consisting of two transmembrane helices without an extracytoplasmic domain. While the cytoplasmic domains of HKs have been studied in detail, the intra-membrane sensing domain systems are still uncharacterized due to difficulties in handling the transmembrane domain. Here, we successfully obtained pure recombinant transmembrane domain of YbdK (YbdK-TM) from E. coli and analyzed the characteristics of YbdK-TM using nuclear magnetic resonance (NMR) and other biophysical methods. YbdK-TM was found to form homo-dimers in DPC micelles based on cross-linking assays and analytical ultracentrifugation analyses. We estimated the size of the YbdK-TM DPC complex to be 46kDa using solution state NMR T(1)/T(2) relaxation analyses in DPC micelles. These results provide information that will allow functional and structural studies of intra-membrane sensing HKs to begin.
authors.affiliation: Division of Magnetic Resonance, Korea Basic Science Institute (KBSI), 804-1, Yangcheong-Ri, Ochang, Chungbuk 363-883, Republic of Korea.
authors.forename: Young Pil
authors.initials: YP
authors.lastname: Kim
authors.forename: Kwon Joo
authors.initials: KJ
authors.lastname: Yeo
authors.forename: Myung Hee
authors.initials: MH
authors.lastname: Kim
authors.forename: Young-Chang
authors.initials: YC
authors.lastname: Kim
authors.forename: Young Ho
authors.initials: YH
authors.lastname: Jeon
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D002462:Cell Membrane / Q000201:enzymology*; D000071677:Histidine Kinase; D008823:Micelles; D019906:Nuclear Magnetic Resonance, Biomolecular; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000737:chemistry; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics; D017434:Protein Structure, Tertiary
chemical_list: D008823:Micelles; D010767:Phosphorylcholine; C028810:dodecylphosphocholine; D011494:Protein Kinases; D000071677:Histidine Kinase