Nature 341, 462 - 464 (05 October 1989); doi:10.1038/341462a0

The lytic protease pro-region does not require a physical linkage to activate the protease domain in vivo

Joy L. Silen & David A. Agard

Department of Biochemistry and Biophysics, and The Howard Hughes Medical Institute, University of California, San Francisco, California 94143, USA

-LYTlC protease, an extracellular serine protease of Lysobacter enzymogenes 495 , is synthesized as a pre-pro-protein¹. Previously it has been shown that when expressed in Escherichia coli, the protein is autocatalytically processed in the periplasmic space, and that the functional protease domain accumulates extracel-lularly². Engineered proteins lacking the 166 amino-acid pro-region were enzymatically inactive and remained cell-associated². By independently expressing the pro- and protease domains in vivo, evidence is provided here that direct covalent linkage is not required for production of active protease. We postulate that the pro-region acts as a template to promote the folding of the protease domain into an active configuration. Our results, combined with recent experiments on the evolutionarily unrelated subtilisin E (ref. 3), suggest that the ability of the pro-region of these bacterial proteases to facilitate folding of their protease domains is not a curiosity of a single system, but may reflect a general property of extracellular bacterial serine proteases.

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