Sequestration and turnover of guinea-pig milk proteins and chicken ovalbumin in Xenopus oocytes.
The stability and distribution of proteins within the living cell can be studied using Xenopus laevis oocytes. Microinjection of messenger RNAs and secretory proteins, followed by cell fractionation, shows that transfer of ovalbumin and milk proteins across intracellular membranes of the oocyte only occurs during their synthesis. Thus milk protein primary translation products, made in the wheat germ cell-free system, when injected into oocytes remain in the cytosol and are not recovered within membrane vesicles. Such miscompartmentalized primary milk proteins are rapidly degraded (t 1/2 0.6 +/- 0.1 h). In contrast, processed milk proteins, extracted from oocytes injected with mammary gland RNA, are relatively stable when introduced into the cytosolic compartment (t 1/2 alpha-lactalbumin 20 +/- 8 h, casein A 6 h, casein B 4 h, casein C 8.3 h). The primary ovalbumin product is also stable (t 1/2 22 +/- 9 h). Indirect evidence that rapid degradation of miscompartmentalized milk protein primary translation products may occur in vivo was obtained by the injection of massive amounts of ovalbumin and milk protein mRNA. Under these conditions there is no accumulation of primary milk protein translation products, but a polypeptide resembling the unglycosylated ovalbium wheat germ primary product can be detected in the cytosol. Only the glyclosylated forms of ovalbumin are found in the oocyte membrane vesicle fraction. We discuss the roles played by the presence of detachable signal sequences and the absence of secondary modifications in determining the rate of degradation of primary translation products within the cytosol.
D000818:Animals; D002645:Chickens; D005260:Female; D006168:Guinea Pigs; D008894:Milk Proteins / Q000378:metabolism*; D008970:Molecular Weight; D009865:Oocytes / Q000378:metabolism*; D010047:Ovalbumin / Q000378:metabolism*; D010063:Ovum / Q000378:metabolism*; D010944:Plants / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D014908:Triticum / Q000378:metabolism; D014981:Xenopus
D008894:Milk Proteins; D012333:RNA, Messenger; D010047:Ovalbumin