Resonance assignments for Oncostatin M, a 24-kDa α-helical protein

Ross C. Hoffman, Franklin J. Moy, Virginia Price, Jane Richardson, Dennis Kaubisch, Eric A. Frieden, Jonathan D. Krakover, Beverly J. Castner, Julie King, Carl J. March, Robert Powers

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

Oncostatin M (OM) is a cytokine that shares a structural and functional relationship with interleukin-6, leukemia inhibitory factor, and granulocyte-colony stimulating factor, which regulate the proliferation and differentiation of a variety of cell types. A mutant version of human OM in which two N-linked glycosylation sites and an unpaired cysteine have been mutated to alanine (N76A/C81A/N193A) has been expressed and shown to be active. The triple mutant has been doubly isotope-labeled with 13C and 15N in order to utilize heteronuclear multidimensional NMR techniques for structure determination. Approximately 90% of the backbone resonances were assigned from a combination of triple-resonance data (HNCA, HNCO, CBCACONH, HBHACONH, HNHA and HCACO), intraresidue and sequential NOEs (3D 15N-NOESY-HMQC and 13C-HSQC-NOESY) and side-chain information obtained from the CCONH and HCCONH experiments. Preliminary analysis of the NOE pattern in the 15N-NOESY-HMQC spectrum and the 13Cα secondary chemical shifts predicts a secondary structure for OM consisting of four α-helices with three intervening helical regions, consistent with the four-helix-bundle motif found for this cytokine family. As a 203-residue protein with a molecular weight of 24 kDa, Oncostatin M is the largest α-helical protein yet assigned.

Original languageEnglish (US)
Pages (from-to)273-282
Number of pages10
JournalJournal of Biomolecular NMR
Volume7
Issue number4
StatePublished - 1996

Keywords

  • Multidimensional NMR
  • Oncostatin M
  • Resonance assignments
  • Triple-resonance spectroscopy

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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    Hoffman, R. C., Moy, F. J., Price, V., Richardson, J., Kaubisch, D., Frieden, E. A., Krakover, J. D., Castner, B. J., King, J., March, C. J., & Powers, R. (1996). Resonance assignments for Oncostatin M, a 24-kDa α-helical protein. Journal of Biomolecular NMR, 7(4), 273-282.