Insights into topology and membrane interaction characteristics of plastoglobule-localized AtFBN1a and AtLOX2.

Plant chloroplasts harbor ubiquitous lipid droplets called plastoglobules. While physically connected to the thylakoid membrane, they are characterized by a unique set of about 30 proteins specifically associated with the plastoglobule. How these proteins selectively target the plastoglobule remains unknown. Protease shaving assays with isolated Arabidopsis thaliana thylakoid and plastoglobule show that a ca. 25 kD portion of the abundant structural protein of plastoglobules, Fibrillin 1a, is protected from protease digestion. Mapping of protease cleavage sites and experimentally identified phosphorylation sites onto a homology model of Fibrillin 1a indicates that this protected sequence corresponds to the C-terminal lipocalin-like domain, implicated in specific lipid binding. In contrast, protease shaving and membrane washing assays with another plastoglobule-associated protein harboring a C-terminal PLAT domain, Lipoxygenase 2, is consistent with an exposed PLAT domain positioned parallel with, and upon, the surface of the plastoglobule. We propose a model where conserved lipid-binding domains associate with either the surface or neutral core of the lipid droplet. Our study provides insight into the topology and membrane interactions of two plastoglobule-localized proteins.