THE IN VITRO INCORPORATION OF C14-GLYCINE INTO ANTIBODY AND OTHER PROTEIN FRACTIONS BY POPLITEAL LYMPH NODES OF RABBITS FOLLOWING THE LOCAL INJECTION OF CRYSTALLINE OVALBUMIN
1956 Volume 43 Issue 5 Pages 653-668
Details
1956 Volume 43 Issue 5 Pages 653-668
Following the injection of crystalline ovalbumin into the foot-pads of rabbits, the popliteal lymph nodes were removed and the incorporation of 1-C14-glycine into antibody and other cellular protein fractions was studied, using cell suspensions or cell free homogenates.
The following results were obtained.
1. Under suitable aerobic conditions the cell suspensions of the lymph nodes from immunized rabbits incorporated very rapidly C14-glycine into antibody. The rates of incorporation of glycine into antibody, soluble protein, the protein moiety of ribonucleoprotein and that of desoxyribonucleoprotein, expressed as μM glycine per g. protein per hour, were 11.9, 3.5, 2.1, and 0.9, respectively, when glycine concentration was 0.68mM.
2. The rate of the C14 glycine uptake into antibody was higher than the proteins of the cellular components, fractionated by the procedure of Schneider, of which that of the microsome fraction showed the highest value, that of the soluble fraction the next, and protein moiety of desoxyribonucleoprotein the lowest value.
3. Aresenate, azide, dinitrophenol, monoiodoacetate and anaerobiosis inhibited the incorporation of glycine into antibody as well as into other cellular proteins. Under anaerobic conditions, the addition of glucose, ATP, and other P-compounds restored only slightly the C14-glycine uptake.
4. When the cell free homogenate was used, the rate of the incorporation was greatly depressed. But even in this condition, the isotope uptake into antibody was more rapid than that into other protein fractions. The arsenate, monoiodoacetate, 2.4-dinitrophenol, or malonate inhibited the amino acid uptake in the cell free homogenate system.
This work was supported by a Grant in Aid for Scientific Research from the Department of Education.
The authors express sincerely their gratitude to Prof. Masuo Ogata and Issei Otawara for their advice and encouragement during this work.
