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THE PLANT CELL, Vol 5, Issue 11 1651-1659, Copyright © 1993 by American Society of Plant Biologists
Molecular Characterization of a Vacuolar Processing Enzyme Related to a Putative Cysteine Proteinase of Schistosoma mansoni
I. Hara-Nishimura, Y. Takeuchi and M. Nishimura
Department of Cell Biology, National Institute for Basic Biology, Okazaki 444, Japan
Proproteins of various vacuolar proteins are post-translationally processed
into mature forms by the action of a unique vacuolar processing enzyme. If
such a processing enzyme is transported to vacuoles together with
proprotein substrates, the enzyme must be a latent form. Immunocytochemical
localization of a vacuolar processing enzyme, a 37-kD cysteine proteinase,
in the endosperm of maturing castor bean seeds places the enzyme in the
vacuolar matrix, where a variety of proproteins is also present. To
characterize a molecular structure of vacuolar processing enzyme, we
isolated a cDNA for the enzyme. Deduced primary structure of a 55-kD
precursor is 33% identical to a putative cysteine proteinase of the human
parasite Schistosoma mansoni. The precursor is composed of a signal
peptide, a 37-kD active processing enzyme domain, and a propeptide
fragment. Although the precursor expressed in Escherichia coli has no
vacuolar processing activity, a 36-kD immunopositive protein expressed in
E. coli is active. These results suggest that the activation of the
vacuolar processing enzyme requires proteolytic cleavage of a 14-kD
C-terminal propeptide fragment of the precursor.
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