Phosphatidylcholine is transferred from chemically-defined liposomes to chloroplasts through proteins of the chloroplast outer envelope membrane

•Chloroplasts take up PC from unilamellar liposomes in a time and temperature dependent fashion.•ATP and GTP are dispensable for in vitro PC uptake into chloroplasts.•Envelope proteins susceptible to thermolysin facilitate PC uptake to chloroplasts.•Increased liposome radius accelerates PC uptake to chloroplasts.

Chloroplasts maintain their lipid balance through a tight interplay with the endoplasmic reticulum (ER). The outer envelope membrane of chloroplasts contains a large proportion of the phospholipid phosphatidylcholine (PC), which is synthesized in the ER and also a possible precursor for thylakoid galactolipids. The mechanism for PC transport from the ER to chloroplasts is not known. Using isolated chloroplasts and liposomes containing radiolabeled PC we investigated non-vesicular transport of PC in vitro. PC uptake in chloroplasts was time and temperature dependent, but nucleotide independent. Increased radius of liposomes stimulated PC uptake, and protease treatment of the chloroplasts impaired PC uptake. This implies that the chloroplast outer envelopes contains an exposed proteinaceous machinery for the uptake of PC from closely apposed membranes.