Structural characterization of a murine myeloid leukaemia inhibitory factor

Abstract

A leukemia inhibitory factor (LIF) which induces macrophage differentiation in M1 murine myeloid leukemia cells and suppresses their proliferation in vitro has been isolated in sufficient quantities (30 .mu.g) from Krebs ascites tumour cell conditioned medium to permit its partial characterization by amino acid sequence analysis. The combination of sensitive microbore column (1.0 and 2.1 mm internal diameter) HPLC technology and microsequence analysis has enabled the positive identification of 125 of the total 179 amino acid residues (70%) in the molecule. The amino acid sequence data reported here permitted the isolation of a partial cDNA clone encoding LIF [Gearing et al. (1978) EMBO J. 6, 3995-4002]. A candidate C-terminus of the LIF molecule predicted from the amino acid sequence was confirmed by subsequent isolation of a cDNA clone corresponding to the C-terminus of the protein. No strong similarity was revealed when the amino acid sequence of LIF was compared with other haemopoietic growth factors, in particular granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor and tumour necrosis factor .alpha. or interleukins. The protein sequence data reported here indicate three sites of post-translational modification (N-linked glycosylation).