J. Willems et al., PURIFICATION AND SEQUENCE OF RAT EXTRACELLULAR-SUPEROXIDE DISMUTASE-BSECRETED BY C(6) GLIOMA, The Journal of biological chemistry, 268(33), 1993, pp. 24614-24621
An enzyme which converts radical oxygen, produced by phorbol 12-myrist
ate 13-acetate activated neutrophils, into nonluminescent products is
secreted by rat C6 glioma. The enzyme was purified from chemically def
ined conditioned media and identified as an extracellular superoxide d
ismutase (EC-SOD). The purified enzyme is distinct from human EC-SOD C
(Hjalmarsson, K., Marklund, S. L., Engstrom, A., and Edlund, T. (1987
) Proc. Natl. Acad. Sci. U. S. A. 84, 6340-6344) by its elution from h
eparin-Sepharose at 300-400 mM NaCl, its pI of 6.1-7.2, and its native
M(r) of 85,000 +/- 20,000. The rat EC-SOD is a dimer with a subunit M
(r) of 34,000-36,000 and is extensively modified by post-translational
processing. Although rat EC-SOD has a high sequence homology with the
catalytic center and the polybasic heparin-binding site near the COOH
terminus of human EC-SOD C, its NH2-terminal sequence and the sequenc
es flanking the heparin-binding site differ substantially. The sequenc
e of the isolated rat EC-SOD cDNA fully confirms the data obtained fro
m amino acid sequence analysis. The amino acid sequence of the enzyme
and its biochemical properties support its identification as the rat E
C-SOD B.