The specificity of glycolipid-preferredoxin interaction: requirements for membrane binding

Abstract

Preferredoxin (prefd) is a precursor protein that is imported into chloroplasts. Monolayer experiments have shown that prefd has a high affinity for monogalactosyldiglyceride (MGaIDG) isolated from chloroplasts, which contains polyunsaturated fatty acid constituents and is therefore in a liquid-expanded state, but has been found to interact also with MGaIDG with long-chain saturated fatty acids, which exist in a gel state. For an optimal interaction, the fatty acid chain length and the extent of unsaturation are also important parameters, whereas the conformation of the sugar moiety, the sugar-glycerol or glycerol-hydrocarbon chain linkages are of little influence on the pressure changes measured in monomolecular layers. Conversely, steric hindrance of a methyl group at position 3 of the sugar largely inhibits the interaction. Quantification of the interaction with radiolabelled prefd shows that only a small part of the molecule is able to penetrate MGaIDG in the gel state, whereas a nearly four-times larger part is able to penetrate MGaIDG isolated from chloroplasts. It is likely that interactions of the transit sequence of prefd with the glycolytic head group MGaIDG are involved in targeting and binding to the chloroplast membrane.

Key Words:

• Protein import
• preferredoxin
• chloroplast
• mono-molecular layer
• glycolipid