Study of lipid-protein interaction in the secretary pathway of plant cell by raising and using antilipid antibodies against particular lipids and proteins in Arabidopsis and tobacco plant

摘要

The lipids and proteins are transported through a well developed system between endoplasmic reticulum (ER) and Golgi and then to all other parts in the Eukaryotes. Lipid and protein based machineries for proteins transport are used in the secretory pathway that are rather well known in animal and yeast cells but far less in plant cells. SNAREs are proteins that interact with others lipids and proteins and are expected to be key regulators of membrane trafficking between ER and the Golgi. SNAREs proteins and their interaction with lipids such as acyl-CoAs was studied and found their presence and involvement in membrane fusion events in the secretory pathway of Arabidopsis comparing with tobacco plant. We produced recombinant proteins for 3 SNAREs and prepared antibodies against them. We purified, anti memb 11, anti Sec22 and anti sed5 IgGs from the sera and tested them in Arabidopsis to see if native proteins would be recognized. We had also prepared anti acyl-CoAs and tested and found that they can detect the acyl-CoAs when they are in interaction with some proteins, so these antibodies could help us to observe proteins interacting with acyl-CoAs. In fact, we used anti acyl-CoAs antibodies in immunoblot and found that they labeled some proteins in Arabidopsis and tobacco. One of these proteins was Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH).