Abstract
We performed the proteomic analysis of the Golgi membranes in rice cell to clarify the structure and function of rice Golgi complex. We isolated the IDPase-associated Golgi membranes by a discontinuous sucrose density gradient centrifugation. The proteins existed in this fraction were analyzed by 2-D electrophoresis-MS and shot gun analyses. We wre able to detect seventy well-known Golgi-localized proteins such as N-acetylglucosaminyltransferase Ι-like protein in the fraction. But many ptoteins other than Golgi-localized were also identified. In order to obtain the further purified Golgi membranes,rice cells were transformed with 35S::AtSYP-31(cis-Golgi marker)/GFP,and then the AtSYP-31/GFP-labeled Golgi membranes were separated by floating in a sucrose density gradient. The ratio of GFP/total proteins of the Golgi membranes was found to increase 20-fold compared with that of the original micorosomal membranes. The identification of proteins associated with the AtSYP-31/GFP-labeled Golgi membranes will be reported.
