title: Expression and characterization of the integral membrane domain of bacterial histidine kinase SCO3062 for structural studies.
abstract: Bacterial histidine kinases play an important role in the response to external stimuli. Structural studies of the histidine kinase transmembrane domain are challenging due to difficulties in protein expression and sample preparation. After carrying out expression screening of a series of histidine kinases, we investigated sample preparation methods for obtaining high quality samples of the periplasmic and transmembrane domain (PTD) of the bacterial histidine kinase SCO3062. Various sample conditions were tested for their ability to give homogeneous NMR spectra of the SCO3062 PTD with well-resolved resonances. Circular dichroism and 3D (15)N-edited NOESY spectrum results demonstrate that the SCO3062 PTD is predominantly alpha-helical. This method should be applicable to the NMR analysis of other transmembrane proteins.
authors.affiliation: Center for Magnetic Resonance, Korea Basic Science Institute, 804-1, Yangcheong-Ri, Ochang, Chungbuk 363-883, Republic of Korea.
authors.forename: Kwon Joo
authors.initials: KJ
authors.lastname: Yeo
authors.forename: Su-Nam
authors.initials: SN
authors.lastname: Kwak
authors.forename: Hyun Jung
authors.initials: HJ
authors.lastname: Kim
authors.forename: Chaejoon
authors.initials: C
authors.lastname: Cheong
authors.forename: Myung Hee
authors.initials: MH
authors.lastname: Kim
authors.forename: Young Ho
authors.initials: YH
authors.lastname: Jeon
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D002462:Cell Membrane / Q000201:enzymology; D002942:Circular Dichroism; D003001:Cloning, Molecular; D005822:Genetic Vectors; D000071677:Histidine Kinase; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D011494:Protein Kinases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; D000071677:Histidine Kinase