Immunolocalization of long-chain acyl-CoAs in plant cells.
Long chain acyl-Coenzyme A esters (acyl-CoAs) are key substrates in many enzymic reactions of lipid metabolism. Due to their amphiphilic nature, the membrane localization of these molecules cannot be established by subcellular membrane fractionation and usual biochemical studies. We have developed another approach based on ultrastructural immunogold cytochemistry. To preserve the acyl-CoA membrane content, the plant material was freeze substituted and cryoembedded after short aldehyde fixation followed by quick freezing. Using Arabidopsis thaliana root cells and specific antibodies raised against acyl-CoAs, we show that acyl-CoAs are mainly localized in endoplasmic reticulum membranes. Our results demonstrate the value of cryo-methods for the accurate localization of labile metabolites in plant cells.
Diakou
Paraskevi
P
Laboratoire de Biogenèse Membranaire, Université Victor Segalen Bordeaux 2, CNRS-UMR 5544, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France.
Fedou
Sandrine
S
Carde
Jean-Pierre
JP
Cassagne
Claude
C
Moreau
Patrick
P
Maneta-Peyret
Lilly
L
D000214:Acyl Coenzyme A / Q000378:metabolism*; D017360:Arabidopsis / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D018517:Plant Roots / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure
D000214:Acyl Coenzyme A