期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1980
卷号：77
期号：3
页码：1556-1560
DOI：10.1073/pnas.77.3.1556
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Proteins labeled with either 3H by reductive methylation or 125I by the chloramine-T method were incubated with Xenopus laevis oocytes; the incorporation and acid precipitability of the proteins were then studied. The uptake rates of both specifically incorporated (vitellogenin) and nonspecifically incorporated proteins (bovine serum albumin and X. laevis serum proteins lacking albumin) were not influenced by the method of labeling. However, 125I-labeled proteins were apparently degraded at rates far exceeding their 3H-labeled counterparts, based on the generation of acid-soluble radioactivity. Thus, after a 3-hr incubation, 3-5 times more 125I-labeled bovine serum albumin and X. laevis serum proteins lacking albumin were degraded than the corresponding 3H-labeled proteins (95% compared to 30% and 75% compared to 15%, respectively), whereas after a 24-hr incubation, the degradation of 125I-labeled vitellogenin was 15 times greater than that of [3H]vitellogenin labeled in vivo (60% compared to 4%). Moreover, examination of the relative amounts of 3H- compared to 125I-labeled bovine serum albumin deposited into the exogenously derived yolk platelet compartment of the oocyte revealed 7 times more acid-precipitable 3H-labeled protein, indicating that the observed discrepancies were not due to reincorporation of the 3H-labeled ligands. Passage of dissolved oocytes previously exposed to 125I-labeled bovine serum albumin (chloramine-T method) over a column of Bio-Gel P-10 revealed some breakdown of bovine serum albumin to intermediate molecular weight components and the presence of a large amount ({approx}90%) of labeled low molecular weight compounds, which analysis showed to be 72% free iodine. The evolution of either iodotyrosine or free iodine would nevertheless be perceived as protein degradation by most analytical procedures (e.g., acid precipitation or autoradiography). We conclude, therefore, that apparent degradation rates observed for endocytotically incorporated proteins may vary depending on the method used to label the protein and caution should be exercised when interpreting results obtained with labeled, particularly chloramine-T labeled, proteins.
关键词：oocyte ; fluid endocytosis ; absorptive endocytosis ; protein turnover
