Reconstitution of phosphatidylserine import into rat liver mitochondria.
The synthesis translocation and decarboxylation of phosphatidylserine occurs in a cell-free system. The principal membrane components necessary are microsomes (source of phosphatidylserine synthase) and mitochondria (source of phosphatidylserine decarboxylase). The interorganelle translocation of phosphatidylserine can be measured by quantitating the decarboxylation of phosphatidyl[1'-14C]serine initially present in prelabeled microsomal membranes using a 14CO2 trapping assay. The decarboxylation of microsomal phosphatidylserine by intact mitochondria is 1) dependent upon substrate (microsomal membrane) concentration, 2) different from decarboxylation of liposomal phosphatidylserine, 3) resistant to proteases, 4) independent of soluble factors, and 5) unaffected by the addition of partially purified phospholipid exchange proteins but accelerated by purified nonspecific phospholipid exchange protein. The rate-limiting step in the reconstituted translocation-decarboxylation system is not the decarboxylation reaction but the initial translocation event between the microsomal membrane and the outer mitochondrial membrane. These data are interpreted to demonstrate that phosphatidylserine import into the mitochondria can occur via collision complexes formed between the endoplasmic reticulum or vesicles derived therefrom and the outer mitochondrial membrane.
Voelker
D R
DR
Lord and Taylor Laboratory for Lung Biochemistry, Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
D000818:Animals; D001692:Biological Transport; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D002262:Carboxy-Lyases / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002474:Cell-Free System; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins; D008862:Microsomes, Liver / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000494:pharmacology; D010718:Phosphatidylserines / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins; D051381:Rats; D011919:Rats, Inbred Strains
D002352:Carrier Proteins; D008081:Liposomes; D008565:Membrane Proteins; D010718:Phosphatidylserines; D049294:Phospholipid Transfer Proteins; D002245:Carbon Dioxide; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase; D010447:Peptide Hydrolases; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase