title: NMR studies of a murine-human chimera of leukaemia inhibitory factor (LIF). Comparison with human LIF.
abstract: Leukaemia inhibitory factor (LIF) is a polyfunctional cytokine active on many cell types. We present here 1H NMR studies on the solution properties and stability of MH35, a chimera of murine and human LIF which can be expressed at high levels in Escherichia coli, thus enabling efficient labelling of the protein with the stable isotopes 13C and 15N. MH35 has 85% sequence identity with human LIF and similar activity in biological assays. The 1H chemical shifts of the 12 conserved aromatic residues and the pKa values of the five conserved histidine residues in MH35 and human LIF are very similar. Temperature dependence studies indicate that both proteins are stable, with significant conformational changes occurring only above 70 degrees C. These results show that these proteins have a similar overall structure and stability and that MH35 is therefore a suitable analogue of human LIF for structural studies in solution.
authors.affiliation: NMR Laboratory, Biomolecular Research Institute, Parkville, Victoria, Australia.
authors.forename: T
authors.initials: T
authors.lastname: Maurer
authors.forename: D K
authors.initials: DK
authors.lastname: Smith
authors.forename: C M
authors.initials: CM
authors.lastname: Owczarek
authors.forename: M J
authors.initials: MJ
authors.lastname: Layton
authors.forename: J G
authors.initials: JG
authors.lastname: Zhang
authors.forename: N A
authors.initials: NA
authors.lastname: Nicola
authors.forename: R S
authors.initials: RS
authors.lastname: Norton
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006131:Growth Inhibitors / Q000737:chemistry*; D006801:Humans; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
chemical_list: D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D008222:Lymphokines; D011993:Recombinant Fusion Proteins