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Purification of a Low Molecular Weight Factor That Induces Differentiation and Inhibits Growth in Myeloid Leukemia Cells¹

School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142 [K. N., N. K., Y. N.], and Institute of Microbial Chemistry, 3-14-23 Kamiosaki, Shinagawa-ku, Tokyo 141 [H. I.], Japan

A procedure is described for purifying a low molecular weight peptide that induces differentiation in mouse myeloid leukemia M1 cells. The factor comes from the conditioned medium of macrophage-like cells differentiated from mouse myeloid leukemia M1 cells. The procedure for purification includes gel filtration on Sephadex G-15, anionic exchange chromatography, thin-layer chromatography, reverse-phase high-performance liquid chromatography on a C₁₈ hydrophobic support, and high-performance liquid chromatography gel filtration. The molecular weight of the factor estimated from the amino acid composition was approximately 1280, which agrees well with that obtained by high-performance liquid chromatography gel filtration. The half-maximal concentration of the purified factor for inducing differentiation of M1 cells was approximately 3.2 x 10^-7 M as judged by nitroblue tetrazolium staining ability. The purified factor also inhibits the growth of human leukemia HL-60 cells.

¹ This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan.

² To whom requests for reprints should be addressed.

Received 1/27/87. Revised 10/29/87. Revised 2/13/88. Revised 4/20/88. Accepted 4/25/88.