The Heparin-binding Domain of Extracellular Superoxide Dismutase Is Proteolytically Processed Intracellularly during Biosynthesis

• Published in: The Journal of biological chemistry Vol. 274; no. 21; pp. 14818 - 14822
• Main Authors: Enghild, Jan J., Thøgersen, Ida B., Oury, Tim D., Valnickova, Zuzana, Højrup, Peter, Crapo, James D.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 21.05.1999; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Culture Techniques; Extracellular Space - metabolism; Heparin - pharmacokinetics; Humans; Mice; Protein Splicing; Rats; Superoxide Dismutase - biosynthesis; Superoxide Dismutase - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.21.14818

Extracellular superoxide dismutase (EC-SOD) is the only known extracellular enzyme designed to scavenge the superoxide anion. The purified enzyme exists in two forms when visualized by reduced SDS-polyacrylamide gel electrophoresis: (i) intact EC-SOD (Trp1–Ala222) containing the C-terminal heparin-binding domain and (ii) cleaved EC-SOD (Trp1–Glu209) without the C-terminal heparin-binding domain. The proteolytic event(s) leading to proteolysis at Glu209–Arg210 and removal of the heparin-binding domain are not known, but may represent an important regulatory mechanism. Removal of the heparin-binding domain affects both the affinity of EC-SOD for and its distribution to the extracellular matrix, in which it is secreted. During the purification of human EC-SOD, the intact/cleaved ratio remains constant, suggesting that proteolytic removal of the heparin-binding domain does not occur during purification (Oury, T. D., Crapo, J. D., Valnickova, Z., and Enghild, J. J. (1996) Biochem. J. 317, 51–57). This was supported by the finding that fresh mouse tissue contains both intact and cleaved EC-SOD. To study other possible mechanisms leading to the formation of cleaved EC-SOD, we examined biosynthesis in cultured rat L2 epithelial-like cells using a pulse-chase protocol. The results of these studies suggest that the heparin-binding domain is removed intracellularly just prior to secretion. In addition, the intact/cleaved EC-SOD ratio appears to be tissue-dependent, implying that the intracellular processing event is regulated in a tissue-specific manner. The existence of this intracellular processing pathway may thus represent a novel regulatory pathway for affecting the distribution and effect of EC-SOD.