Partial disassembly of peroxisomes.
Rat liver peroxisomes were subjected to a variety of procedures intended to partially disassemble or damage them; the effects were analyzed by recentrifugation into sucrose gradients, enzyme analyses, electron microscopy, and SDS PAGE. Freezing and thawing or mild sonication released some matrix proteins and produced apparently intact peroxisomal "ghosts" with crystalloid cores and some fuzzy fibrillar content. Vigorous sonication broke open the peroxisomes but the membranes remained associated with cores and fibrillar and amorphous matrix material. The density of both ghosts and more severely damaged peroxisomes was approximately 1.23. Pyrophosphate (pH 9) treatment solubilized the fibrillar content, yielding ghosts that were empty except for cores. Some matrix proteins such as catalase and thiolase readily leak from peroxisomes. Other proteins were identified that remain in mechanically damaged peroxisomes but are neither core nor membrane proteins because they can be released by pyrophosphate treatment. These constitute a class of poorly soluble matrix proteins that appear to correspond to the fibrillar material observed morphologically. All of the peroxisomal beta-oxidation enzymes are located in the matrix, but they vary greatly in how easily they leak out. Palmitoyl coenzyme A synthetase is in the membrane, based on its co-distribution with the 22-kilodalton integral membrane polypeptide.
Alexson
S E
SE
Fujiki
Y
Y
Shio
H
H
Lazarow
P B
PB
D000818:Animals; D002451:Cell Compartmentation; D002458:Cell Fractionation / Q000379:methods; D011756:Diphosphates; D005260:Female; D005615:Freezing; D008099:Liver / Q000201:enzymology / Q000648:ultrastructure; D008830:Microbodies / Q000201:enzymology / Q000648:ultrastructure*; D008854:Microscopy, Electron; D011092:Polyethylene Glycols; D011506:Proteins / Q000032:analysis; D051381:Rats; D012995:Solubility; D013010:Sonication
D011756:Diphosphates; D011506:Proteins; D011092:Polyethylene Glycols