Analysis of zeins' ER retention in Xenopus oocytes |
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Authors: | Dong-Hee Lee O-Yu Kwon Karl Pedersen |
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Institution: | aDepartment of Biology, 2123 Derring Hall, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061-0406, U.S.A.;bDivision of Endocrinology and Metabolism, Harvard Medical School and Beth Israel Hospital, 99 Brookline Avenue, Boston, MA 02215, U.S.A. |
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Abstract: | Zeins, maize storage proteins, are retained in the endoplasmic reticulum (ER) during the intracellular protein targeting process. Hydrophobic interaction has been postulated as the driving force of zeins' aggregation and retention in the ER. Recently, a class of zein (the 27K zein) has been proposed to facilitate zeins' ER retention by anchoring to the ER membrane. This study investigated the significance of the two proposed mechanisms toward zeins' ER retention using Xenopus oocyte. Following injection of the total or 27K zein mRNA, zein's movement within the ER was analyzed based upon the extent of diffusion to the non-injected oocyte half. This study indicates that the total zeins freely move within the lumen of the ER, thus, suggesting that the intermolecular aggregation, leading to insolubility and exclusion from the ER lumenal fluid, may not be essential for zeins' ER retention. This study also suggests that the 27K zein may not facilitate zeins' ER retention by virtue of an anchor to the ER membrane based on its free movement in the ER. Free movement of the total and 27K zeins, under conditions where zein aggregates should form, necessitates a reevaluation of the mechanisms responsible for zein polypeptides' ER retention and protein body formation. |
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