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Title :Isolation and proteomic analysis of rice Golgi membranes: cis-Golgi membranes labeled with GFP-SYP31
Authors :Asakura, Tsuyoshi
Hirose, Shota
Katamine, Hiroki
Kitajima, Aya
Hori, Hidetaka
Sato, Masa
Fujiwara, Masayuki
Shimamoto, Ko
Mitsui,Toshiaki
Publisher :日本植物細胞分子生物学会
Issue Date :Dec-2006
Journal Title :Plant Biotechnology
Volume :23
Issue :5
Start Page :475
End Page :485
ISSN :1342-4580
Abstract :The Golgi membranes labeled with cis-Golgi marker GFP-SYP31 were isolated from suspension-cultured cells of rice transformed with 35S::GFP-SYP31 by floating through a discontinuous sucrose density gradient in the presence of 5 mM MgCl2. The specific fluorescence intensity of final membrane preparation increased to approximately 150-fold in comparison with that of the post-nucleus soluble fraction. Specific activity of membrane-bound α-mannosidase (cis-Golgi) markedly increased, but NDPase (medial-/trans-Golgi) and NADPH-cytochrome c reductase endoplasmic reticulum were weakly detected in the membrane fraction. The other organelle marker enzymes, cytochrome c oxidase (mitochondria), alkaline pyrophosphatase (plastid), and catalase (peroxisome) were not detectable. Comparative display of protein spots in GFP-SYP31-labeled membranes, microsomal membranes and soluble fraction on the two dimensional gels showed that some proteins are markedably concentrated in the cis-Golgi membrane fraction. Furthermore, the mass spectrometric analysis of proteins separated by SDS-polyacrylamide gel electrophoresis indicated that the highly purified Golgi membranes contained several membrane traffic-related proteins and ER resident proteins. These findings indicated a close relationship between the ER and the cis-Golgi membranes.
Keywords :Cis-Golgi
green fluorescent protein
small GTPase
rice
syntaxins 31
Type Local :学術雑誌論文
Language :eng
Format :application/pdf
URI :http://hdl.handle.net/10191/17594
fullTextURL :http://dspace.lib.niigata-u.ac.jp/dspace/bitstream/10191/17594/1/ZL_23(5)_475-485.pdf
Appears in Collections:10 査読済論文 = Postprint

Please use this identifier to cite or link to this item: http://hdl.handle.net/10191/17594