Elicitins trap and transfer sterols from micelles, liposomes and plant plasma membranes.
Using elicitins, proteins secreted by some phytopathogenic Oomycetes (Phytophthora) known to be able to transfer sterols between phospholipid vesicles, the transfer of sterols between micelles, liposomes and biological membranes was studied. Firstly, a simple fluorometric method to screen the sterol-carrier capacity of proteins, avoiding the preparation of sterol-containing phospholipidic vesicles, is proposed. The transfer of sterols between DHE micelles (donor) and stigmasterol or cholesterol micelles (acceptor) was directly measured, as the increase in DHE fluorescence signal. The results obtained with this rapid and easy method lead to the same conclusions as those previously reported, using fluorescence polarization of a mixture of donor and acceptor phospholipid vesicles, prepared in the presence of different sterols. Therefore, the micelles method can be useful to screen proteins for their sterol carrier activity. Secondly, elicitins are shown to trap sterols from purified plant plasma membranes and to transfer sterols from micelles to these biological membranes. This property should contribute to understand the molecular mechanism involved in sterol uptake by Phytophthora. It opens new perspectives concerning the role of such proteins in plant-microorganism interactions.
Unité Associée INRA/Université de Bourgogne 692, Laboratoire de Phytopharmacie et de Biochimie des Interactions Cellulaires, INRA, BV 1540, 21034, Dijon Cedex, France.
S
S
Vauthrin
V
V
Mikes
M L
ML
Milat
M
M
Ponchet
B
B
Maume
H
H
Osman
J P
JP
Blein
D020418:Algal Proteins; D002352:Carrier Proteins / Q000737:chemistry* / Q000502:physiology; D002462:Cell Membrane / Q000737:chemistry; D002784:Cholesterol; D004875:Ergosterol / Q000031:analogs & derivatives; D005453:Fluorescence; D005656:Fungal Proteins / Q000737:chemistry* / Q000502:physiology; D008081:Liposomes / Q000737:chemistry*; D008823:Micelles; D010940:Plant Proteins; D010944:Plants / Q000737:chemistry*; D011506:Proteins; D013261:Sterols / Q000737:chemistry*; D013265:Stigmasterol; D013997:Time Factors
D020418:Algal Proteins; D002352:Carrier Proteins; D005656:Fungal Proteins; D008081:Liposomes; D008823:Micelles; D010940:Plant Proteins; D011506:Proteins; D013261:Sterols; C061032:cryptogein protein, Phytophthora cryptogea; C085101:elicitin, Phytophthora; C072087:sterol carrier proteins; C010905:dehydroergosterol; D002784:Cholesterol; D013265:Stigmasterol; D004875:Ergosterol