Distribution of an Ankyrin-repeat Protein on the Endoplasmic Reticulum in Arabidopsis

• Published in: Journal of integrative plant biology Vol. 51; no. 2; pp. 140 - 146
• Main Authors: Wei, Liqin, Li, Yan
• Format: Journal Article
• Language: English
• Published: Melbourne, Australia Melbourne, Australia : Blackwell Publishing Asia 01.02.2009; Blackwell Publishing Asia; State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, China
• Subjects: Ankyrin Repeat; ankyrin-repeat protein; Ankyrins - genetics; Ankyrins - metabolism; Arabidopsis; Arabidopsis - genetics; Arabidopsis - metabolism; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Blotting, Western; Cloning, Molecular; DNA, Complementary - genetics; endoplasmic reticulum; Endoplasmic Reticulum - metabolism; Fluorescent Antibody Technique; Protein Structure, Secondary; subcellular localization; Western blotting; ankyrin-repeat protein; western blotting; Arabidopsis; subcellular localization; endoplasmic reticulum
• ISSN: 1672-9072; 1744-7909
• DOI: 10.1111/j.1744-7909.2008.00791.x

There are many ankyrin-repeat proteins in plant cells. However, the distribution and function of these proteins are mostly unclear. By reverse transcription-polymerase chain reaction, a gene encoding an ankyrin-like protein was cloned from Arabidopsis and named AtANK1 (GenBank accession no. NM_120340). The 6-His-tagged AtAnk1-N fusion protein was affinity-purified and its rabbit polyclonal antibody was obtained. Immuno-blotting with the purified anti-AtAnk1-N polyclonal antibody revealed that the relative molecular weight of the AtANK1 protein was about 76 kDa. By immunofluorescence labeling and immuno-gold labeling with the purified anti-AtAnk1-N polyclonal antibody, coupled with confocal and transmission electron microscopy observation, AtANK1 was found to be distributed on the membrane of the endoplasmic reticulum in Arabidopsis cells. Based on these results, we suggested that AtANK1 might be involved in endoplasmic reticulum-related protein localization and sorting in plant cells.