Age-related change in the degradation rate of ovalbumin microinjected into mouse liver parenchymal cells.
Change in the degradation rate of ovalbumin (OVA) microinjected into liver parenchymal cells isolated from mice of various ages was studied. OVA was injected by osmotic lysis of pinosomes and the amount of OVA was determined by immunoblotting using purified antibody to OVA. Cellular activity as judged by rates of protein synthesis and degradation of pulse-labeled proteins was not affected by the injection. To localize injected OVA in the cells, cell extracts were fractionated by differential centrifugation and the amount of OVA and the activity of beta-D-galactosidase as a lysosomal marker enzyme of each fraction were determined. The ovalbumin was mostly found in the soluble fraction, while the beta-D-galactosidase activity was found in the particulate fraction, indicating that the ovalbumin was spread in the cytosol but was not present in the pinosomes or lysosomes. The average half-lives of OVA were 106, 113, and 164 h in the cells from young (3.5-6.5 months), middle-aged (13.5-20.0 months), and old (24.5-30.5 months) mice, respectively. Thus, the half-life of ovalbumin in the cells of senescent mice was about 50% longer than that in the cells of young or middle-aged mice. These results are in good agreement with those of our previous investigation, which showed that the half-life of inactivation of horseradish peroxidase was extended by about 50% in the hepatocytes from old mice (Ishigami and Goto, 1988, Mech. Ageing Dev., 46, 125-133).
Ishigami
A
A
Department of Biochemistry, School of Pharmaceutical Sciences, Toho University, Chiba, Japan.
Goto
S
S
D000375:Aging; D000818:Animals; D002478:Cells, Cultured; D015151:Immunoblotting; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008845:Microinjections; D010047:Ovalbumin / Q000008:administration & dosage / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D010047:Ovalbumin; D001616:beta-Galactosidase