Resonance assignments, secondary structure and topology of leukaemia inhibitory factor in solution
Details
Publication Year 1997-02,Volume 9,Issue #2,Page 113-126
Journal Title
JOURNAL OF BIOMOLECULAR NMR
Publication Type
Journal Article
Abstract
The chemical shift assignments and secondary structure of a murine-human chimera, MH35, of leukaemia inhibitory factor (LIF), a 180-residue protein of molecular mass 20 kDa, have been determined from multidimensional heteronuclear NMR spectra acquired on a uniformly C-13, N-15-labelled sample. Secondary structure elements were defined on the basis of chemical shifts, NH-(CH)-H-alpha coupling constants, medium-range NOEs and the location of slowly exchanging amide protons. The protein contains four alpha-helices, the relative orientations of which were determined on the basis of long-range, interhelical NOEs. The four helices are arranged in an up-up-down-down orientation, as found in other four-helical bundle cytokines. The overall topology of MH35-LIF is similar to thar of the X-ray crystallographic structure for murine LIF [Robinson et al. (1994) Cell, 77, 1101-1116]. Differences between the X-ray structure and the solution structure are evident in the N-terminal tail, where the solution structure has a trans-Pro(17) compared with the cis-Pro(17) found in the crystal structure and the small antiparallel beta-sheet encompassing residues in the N-terminus and CD loop in the crystal structure is less stable.
Publisher
KLUWER ACADEMIC PUBL
Keywords
COLONY-STIMULATING FACTOR; 3-DIMENSIONAL SOLUTION STRUCTURE; HUMAN INTERFERON-GAMMA; HUMAN GROWTH-HORMONE; KREBS ASCITES-CELLS; CRYSTAL-STRUCTURE; HUMAN INTERLEUKIN-4; NMR-SPECTROSCOPY; N-15 NMR; CHEMICAL-SHIFTS
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Creation Date: 1997-02-01 12:00:00
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