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© The Rockefeller University Press, 0021-9525/2000/2/453/ $5.00
The Journal of Cell Biology, Volume 148, Number 3, February 7, 2000 453-464


Original Article

Mass Transport of Proform of a KDEL-tailed Cysteine Proteinase (SH-EP) to Protein Storage Vacuoles by Endoplasmic Reticulum–derived Vesicle Is Involved in Protein Mobilization in Germinating Seeds

Kiminori Toyookaa, Takashi Okamotoa, and Takao Minamikawaa
a Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa 1-1, Hachioji, Tokyo, 192-0397 Japan

Correspondence to: Takashi Okamoto, Department of Biological Sciences, Tokyo Metropolitan University, Minami-osawa, Hachioji, Tokyo, 192-0397 Japan. Tel:+81 426-77-2559 Fax:+81 426-77-2559 E-mail:okamoto-takashi{at}c.metro-u.ac.jp.

A vacuolar cysteine proteinase, designated SH-EP, is expressed in the cotyledon of germinated Vigna mungo seeds and is responsible for the degradation of storage proteins. SH-EP is a characteristic vacuolar proteinase possessing a COOH-terminal endoplasmic reticulum (ER) retention sequence, KDEL. In this work, immunocytochemical analysis of the cotyledon cells of germinated V. mungo seeds was performed using seven kinds of antibodies to identify the intracellular transport pathway of SH-EP from ER to protein storage vacuoles. A proform of SH-EP synthesized in ER accumulated at the edge or middle region of ER where the transport vesicle was formed. The vesicle containing a large amount of proSH-EP, termed KV, budded off from ER, bypassed the Golgi complex, and was sorted to protein storage vacuoles. This massive transport of SH-EP via KV was thought to mediate dynamic protein mobilization in the cotyledon cells of germinated seeds. We discuss the possibilities that the KDEL sequence of KDEL-tailed vacuolar cysteine proteinases function as an accumulation signal at ER, and that the mass transport of the proteinases by ER-derived KV-like vesicle is involved in the protein mobilization of plants.

Key Words: cysteine proteinase, endoplasmic reticulum, intracellular transport, KDEL sequence, protein storage vacuole


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