Analysis of the major integral membrane proteins of peroxisomes from mouse liver.
Two major proteins with subunit molecular masses of 68 and 70 kDa were isolated from the integral membrane protein fraction of peroxisomes purified from mouse liver. The two proteins were shown to be distinct proteins by two criteria: first, immunoblot analysis demonstrated that antisera against the 68 kDa protein did not cross-react with the 70 kDa protein, and vice versa; and second, the partial peptide maps resulting from proteinase digestion of the proteins were different. Immunoblot analyses to test the specificities of the antisera demonstrated that only the expected molecular mass species in purified peroxisomes, and in membranes prepared from these organelles, were recognized; there was no identification of proteins from purified mitochondrial or microsomal fractions. The concentrations of both of these proteins were increased in livers of mice treated with clofibrate, a hypolipidemic drug and peroxisome proliferator, with the effect being greater for the 70 kDa component. The localization of the 68 kDa protein was shown to be completely integral to the peroxisome membrane. Although some 70 kDa protein was integral to the membrane, a significant proportion was released from the membrane by some procedures believed to detach peripheral proteins. The 70 kDa protein was also particularly susceptible to degradation during isolation - in particular, addition of EDTA to media used for isolation of peroxisomes resulted in membranes in which this protein was degraded to smaller immunoreactive fragments. These data have been discussed in relation to the significant clarification which they have provided of the status and characteristics of the major protein components of peroxisomal membranes.
Chen
N H
NH
Division of Science and Technology, Griffith University, Brisbane, Australia.
Crane
D I
DI
Masters
C J
CJ
D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation / Q000379:methods; D002994:Clofibrate / Q000494:pharmacology; D007425:Intracellular Membranes / Q000032:analysis; D008099:Liver / Q000032:analysis*; D008565:Membrane Proteins / Q000032:analysis*; D051379:Mice; D008830:Microbodies / Q000032:analysis* / Q000201:enzymology; D008970:Molecular Weight; D010449:Peptide Mapping; D011092:Polyethylene Glycols; D012995:Solubility
D008565:Membrane Proteins; D011092:Polyethylene Glycols; D002994:Clofibrate