use of polyclonal antibodies to identify peptides exposed on the stroma side of the spinach thylakoid

We have raised polyclonal antibodies against an oxygen-evolving photosystem II preparation. Western Blot analysis of the whole serum revaals antibodies specific for at least 15 Coomassie visible bands ranging from 59 to 11 kDa. These antibodies are specific for proteins located on both sides of the membrane. Included are antibodies specific for Tris-removable peptides (33, 25 and 18 kda), which are thought to be exposed on the lumen surface of the PS II complex. Since the whole serum agglutinates thylakoids, antibodies specific for the stroma side of the PS II complex are also present. A sub-population of antibodies can be isolated by allowing the antibodies in whole serum to bind to EDTA-treated thylakoid membranes. The antibodies which specifically bind are cross-reactive with peptides with Mr of 59, 57, 34, 28, 27, 26, and 23 kDa. Our data indicate that these peptides have antigenic determinants exposed on the stroma side of the thylakoid membrane.