Plant and Cell Physiology, 1995, Vol. 36, No. 8 1599-1606
© 1995
Transient and Specific Expression of a Cysteine Endopeptidase Associated with Autolysis during Differentiation of Zinnia Mesophyll Cells into Tracheary Elements
Biological Institute, Faculty of Science, Tohoku University Aoba-Aramaki, Aoba-ku, Sendai, 980-77 Japan
1To whom correspondence should be addressed.
Endopeptidase activities during the differentiation of Zinnia cells into tracheary elements (TEs) were examined with several peptidyl 4-methyl-7-coumarylamido (MCA) as substrates. The activity that hydrolysed carbobenzoxy-Phe-Arg-MCA (Z-Phe-Arg-MCA) at pH 5 increased in a differentiation-related manner: this activity, which was not observed in freshly isolated mesophyll cells was induced by the combination of 
-naphthaleneacetic acid (NAA) and 6-benzyladenine (BA) that is necessary for differentiation of TEs, but not by NAA or BA alone. The activity in cells cultured in TE-inductive medium that contained both NAA and BA increased very rapidly between the 48th and 60th hours of culture, when the number of TE increased rapidly. A protease responsible for this activity with a molecular mass of 30 kDa was partially purified from cells which had been cultured in the TE-inductive medium and included many immature TEs. Strong inhibition by [L-3-trans-carboxyoxiran-2-carbonyl]-L-Leu-agmatin (E-64), and activation by dithiothreitol (DTT) indicated that this protease belongs to a family of cysteine endopeptidases (EC 3.4.22).
1Present address: Department of Material and Biological Engineering, Tsuruoka National College of Technology, Inooka-Sawada 104, Tsuruoka, Yamagata, 997 Japan
(Received July 11, 1995; Accepted September 29, 1995)
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