Investigating the interaction between the violaxanthin cycle enzyme zeaxanthin epoxidase and the thylakoid membrane

In the present study the interaction between the violaxanthin cycle enzyme zeaxanthin epoxidase (ZEP) and the thylakoid membrane was investigated. Isolated, active thylakoid membranes of spinach (Spinacia oleracea L.) were subjected to different salt and detergent treatments that are generally used to isolate peripheral and integral membrane proteins. These salt and detergent treatments included the use of the salts NaBr, Na2CO3 and Tris and the detergents octylglucoside (OG) and dodecylmaltoside (DM). After the treatments the activity of the ZEP was determined in washed thylakoid membranes. To obtain additional information about the mode of ZEP binding to the membrane a hydrophobicity plot based on the amino acid sequence of the protein was constructed. The plot was then compared to a diagram obtained for the photosystem II antenna Lhcb1 protein whose integration into the thylakoid membrane is known. The results of the salt and detergent treatments of the thylakoid membrane suggest that the ZEP is a peripheral, rather weakly bound membrane protein. Results from the hydrophobicity plots indicate the existence of specialized protein domains which may realize the partial integration and binding of the ZEP to the thylakoid membrane.