Purification of an Inhibitor of Erythroid Progenitor Cell Cycling and Antagonist to Interleukin 3 from Mouse Marrow Cell Supernatants and Its Identification as Cytosolic Superoxide Dismutase

We have isolated a protein from media conditioned by a murine marrow-derived cell line (PB6) and from mouse marrow supernatants that antagonizes interleukin 3-dependent proliferation of cells in culture and reversibly inhibits DNA synthesis of erythroid progenitor cells (BFU-E) in vitro. This protei...

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Bibliographic Details
Published in:The Journal of cell biology 1990-09, Vol.111 (3), p.1217-1223
Main Authors: Pluthero, Fred G., Shreeve, Mona, Eskinazi, Denise, van der Gaag, Henk, Huang, Kuo-Sen, Hulmes, Jeffrey D., Blum, Max, Axelrad, Arthur A.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Bone marrow
Bone Marrow - enzymology
Cell Cycle - physiology
Cell Division - physiology
Cell growth
Cell lines
Cells
Chromatography, Ion Exchange
Enzymes and enzyme inhibitors
Erythroid cells
Erythroid Precursor Cells - cytology
Erythroid progenitor cells
Fundamental and applied biological sciences. Psychology
Gels
Interleukin-3 - antagonists & inhibitors
Mice
Mice, Inbred C57BL
Molecular Sequence Data
Oxidoreductases
Stem cells
Superoxide Dismutase - isolation & purification
Superoxide Dismutase - physiology
Superoxides
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Summary:We have isolated a protein from media conditioned by a murine marrow-derived cell line (PB6) and from mouse marrow supernatants that antagonizes interleukin 3-dependent proliferation of cells in culture and reversibly inhibits DNA synthesis of erythroid progenitor cells (BFU-E) in vitro. This protein, p16 (monomer M r = 16 kD on SDS-PAGE), was purified to homogeneity and amino acid sequencing of a polypeptide fragment yielded a sequence identical to that of murine cytosolic Cu,Zn-containing superoxide dismutase (SOD). The identification of p16 as SOD was confirmed by the detection of SOD enzymatic activity in pure p16 fractions, and when a commercial human erythrocytic SOD preparation was tested it showed the same cell inhibitory activities as p16. These observations show that superoxide dismutase is able to affect the cycling and growth factor responses of hematopoietic cells, activities that have not previously been associated with this enzyme.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.111.3.1217