Abstract
Turnover studies of the surface membrane and of cell particulate matter of L cells in tissue culture in logarithmic and plateau phase of growth have been made. The rate of incorporation of isotope into these fractions and the rate of fall of specific activities of labeled L-cell fractions have been observed. The following interpretation of the data appears most likely although other interpretations are possible. Growing and nongrowing cells synthesize approximately similar amounts of surface membrane and particulate material. In the growing cell the material is incorporated with net increases in substance. There is relatively little turnover. In the nongrowing cell newly synthesized material is incorporated, but a corresponding amount of material is eliminated so that there is turnover without net increase of substance. Our results suggest that there is no gross differential turnover between the protein, lipid, and carbohydrate of the surface membrane under the conditions of our experiments. Metabolic inhibitors or omission of amino acids in the culture medium lead to a decrease in synthesis of surface membrane and cell particulates and cause an equivalent decrease in the rate of degradation of surface membrane and of particulates; therefore the synthetic and degradative aspects of turnover appear to be coupled. As cultures of nongrowing cells in suspension or on a glass surface age, their synthetic and turnover capacities diminish. Our results suggest that the cell may exist in a nongrowing state with a level of synthesis similar to that of a growing cell. It can exist in this state with a high level of turnover.
Full Text
The Full Text of this article is available as a PDF (1.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BALLOU J. E., THOMPSON R. C. Studies of metabolic turnover with tritium as a tracer. V. The predominantly non-dynamic state of body constituents in the rat. J Biol Chem. 1956 Dec;223(2):795–809. [PubMed] [Google Scholar]
- BASERGA R. THE RELATIONSHIP OF THE CELL CYCLE TO TUMOR GROWTH AND CONTROL OF CELL DIVISION: A REVIEW. Cancer Res. 1965 Jun;25:581–595. [PubMed] [Google Scholar]
- Borek E., Ponticorvo L., Rittenberg D. PROTEIN TURNOVER IN MICRO-ORGANISMS. Proc Natl Acad Sci U S A. 1958 May;44(5):369–374. doi: 10.1073/pnas.44.5.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DE GIER J., VAN DEENENL A DIETARY INVESTIGATION ON THE VARIATIONS IN PHOSPHOLIPID CHARACTERISTICS OF RED-CELL MEMBRANES. Biochim Biophys Acta. 1964 Jun 15;84:294–304. doi: 10.1016/0926-6542(64)90057-5. [DOI] [PubMed] [Google Scholar]
- Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. I. Structural and chemical differentiation in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):73–96. doi: 10.1083/jcb.30.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. II. Synthesis of constitutive microsomal enzymes in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):97–117. doi: 10.1083/jcb.30.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
- EAGLE H., PIEZ K. A., FLEISCHMAN R., OYAMA V. I. Protein turnover in mammaliar cell cultures. J Biol Chem. 1959 Mar;234(3):592–597. [PubMed] [Google Scholar]
- EAGLE H., PIEZ K. A., FLEISCHMAN R. The utilization of phenylalanine and tyrosine for protein synthesis by human cells in tissue culture. J Biol Chem. 1957 Oct;228(2):847–861. [PubMed] [Google Scholar]
- EIDAM C. R., MERCHANT D. J. THE PLATEAU PHASE OF GROWTH OF THE L-M STRAIN MOUSE CELL IN A PROTEIN-FREE MEDIUM. 3. THE EFFECT OF ADDED GLUCOSE ON PROTEIN AND NUCLEIC ACID SYNTHESIS AND ON CARBOHYDRATE UTILIZATION. Exp Cell Res. 1965 Jan;37:147–160. doi: 10.1016/0014-4827(65)90165-5. [DOI] [PubMed] [Google Scholar]
- EIDAM C. R., MERCHANT D. J. THE PLATEAU PHASE OF GROWTH OF THE L-M STRAIN MOUSE CELL IN A PROTEIN-FREE MEDIUM. I. PATTERNS OF PROTEIN AND NUCLEIC ACID SYNTHESIS AND TURNOVER. Exp Cell Res. 1965 Jan;37:132–139. doi: 10.1016/0014-4827(65)90163-1. [DOI] [PubMed] [Google Scholar]
- FORSSBERG A., REVESZ L. A study on the metabolic state of proteins in the cells of two ascites tumors. Biochim Biophys Acta. 1957 Jul;25(1):165–171. doi: 10.1016/0006-3002(57)90435-3. [DOI] [PubMed] [Google Scholar]
- GREENLEES J., LEPAGE G. A. Protein turnover in a study of host-tumor relationships. Cancer Res. 1955 May;15(4):256–262. [PubMed] [Google Scholar]
- HAGERMAN J. S., GOULD R. G. The in vitro interchange of cholesterol between plasma and red cells. Proc Soc Exp Biol Med. 1951 Oct;78(1):329–332. doi: 10.3181/00379727-78-19064. [DOI] [PubMed] [Google Scholar]
- HALVORSON H. Intracellular protein and nucleic acid turnover in resting yeast cells. Biochim Biophys Acta. 1958 Feb;27(2):255–266. doi: 10.1016/0006-3002(58)90332-9. [DOI] [PubMed] [Google Scholar]
- HALVORSON H. Studies on protein and nucleic acid turnover in growing cultures of yeast. Biochim Biophys Acta. 1958 Feb;27(2):267–276. doi: 10.1016/0006-3002(58)90333-0. [DOI] [PubMed] [Google Scholar]
- HARRIS H., WATTS J. W. Turnover of protein in a non-multiplying animal cell. Nature. 1958 Jun 7;181(4623):1582–1584. doi: 10.1038/1811582b0. [DOI] [PubMed] [Google Scholar]
- Hirsch C. A., Hiatt H. H. Turnover of liver ribosomes in fed and in fasted rats. J Biol Chem. 1966 Dec 25;241(24):5936–5940. [PubMed] [Google Scholar]
- JORDAN H. C., MILLER L. L., PETERS P. A. Constant protein catabolism of Walker carcinoma 256 and human skin epithelium in tissue culture. Cancer Res. 1959 Feb;19(2):195–200. [PubMed] [Google Scholar]
- JORDAN H. C., SCHMIDT P. A. Constant protein turnover in mammalian cells during logarithmic growth. Biochem Biophys Res Commun. 1961 Mar 24;4:313–316. doi: 10.1016/0006-291x(61)90242-x. [DOI] [PubMed] [Google Scholar]
- KANFER J., KENNEDY E. P. METABOLISM AND FUNCTION OF BACTERIAL LIPIDS. I. METABOLISM OF PHOSPHOLIPIDS IN ESCHERICHIA COLI B. J Biol Chem. 1963 Sep;238:2919–2922. [PubMed] [Google Scholar]
- KING D. W., BENSCH K. G., HILL R. B., Jr State of dynamic equilibrium in protein of mammalian cells. Science. 1960 Jan 8;131(3393):106–107. doi: 10.1126/science.131.3393.106. [DOI] [PubMed] [Google Scholar]
- KOCH A. L., LEVY H. R. Protein turnover in growing cultures of Escherichia coli. J Biol Chem. 1955 Dec;217(2):947–957. [PubMed] [Google Scholar]
- Kornfeld S., Ginsburg V. The metabolism of glucosamine by tissue culture cells. Exp Cell Res. 1966 Mar;41(3):592–600. doi: 10.1016/s0014-4827(66)80109-x. [DOI] [PubMed] [Google Scholar]
- Kruse P. F., Jr, Miedema E., Carter H. C. Amino acid utilizations and protein synthesis at various proliferation rates, population densities, and protein contents of perfused animal cell and tissue culture. Biochemistry. 1967 Apr;6(4):949–955. doi: 10.1021/bi00856a001. [DOI] [PubMed] [Google Scholar]
- LEVINE E. M., BECKER Y., BOONE C. W., EAGLE H. CONTACT INHIBITION, MACROMOLECULAR SYNTHESIS, AND POLYRIBOSOMES IN CULTURED HUMAN DIPLOID FIBROBLASTS. Proc Natl Acad Sci U S A. 1965 Feb;53:350–356. doi: 10.1073/pnas.53.2.350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOVELOCK J. E., JAMES A. T., ROWE C. E. The lipids of whole blood. 2. The exchange of lipids between the cellular constituents and the lipoproteins of human blood. Biochem J. 1960 Jan;74:137–140. doi: 10.1042/bj0740137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- LUCK D. J. Genesis of mitochondria in neurospora crassa. Proc Natl Acad Sci U S A. 1963 Feb 15;49:233–240. doi: 10.1073/pnas.49.2.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MANDELSTAM J. The intracellular turnover of protein and nucleic acids and its role in biochemical differentiation. Bacteriol Rev. 1960 Sep;24(3):289–308. doi: 10.1128/br.24.3.289-308.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MANDELSTAM J. Turnover of protein in growing and non-growing populations of Escherichia coli. Biochem J. 1958 May;69(1):110–119. doi: 10.1042/bj0690110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MARKOVITZ A., KLEIN H. P. On the sources of carbon for the induced biosynthesis of alpha-amylase in Pseudomonas saccharophila. J Bacteriol. 1955 Dec;70(6):649–655. doi: 10.1128/jb.70.6.649-655.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MERCHANT D. J., EIDAM C. R. THE PLATEAU PHASE OF GROWTH OF THE L-M STRAIN MOUSE CELL IN A PROTEIN-FREE MEDIUM. II. PROLONGATION OF THE PLATEAU PHASE BY SUPPLEMENTAL GLUCOSE. Exp Cell Res. 1965 Jan;37:140–146. doi: 10.1016/0014-4827(65)90164-3. [DOI] [PubMed] [Google Scholar]
- MOLDAVE K. Intracellular protein metabolism in Ehrlich's ascites carcinoma cells. J Biol Chem. 1956 Jul;221(1):543–553. [PubMed] [Google Scholar]
- MOLDAVE K. The release of labeled constituents from cellular fractions of Ehrlich ascites cells. J Biol Chem. 1957 Apr;225(2):709–714. [PubMed] [Google Scholar]
- MURPHY J. R. Erythrocyte metabolism. IV. Equilibration of cholesterol-4-C-14 between erythrocytes and variously treated sera. J Lab Clin Med. 1962 Oct;60:571–578. [PubMed] [Google Scholar]
- Omura T., Siekevitz P., Palade G. E. Turnover of constituents of the endoplasmic reticulum membranes of rat hepatocytes. J Biol Chem. 1967 May 25;242(10):2389–2396. [PubMed] [Google Scholar]
- PARK J. T., JOHNSON M. J. A submicrodetermination of glucose. J Biol Chem. 1949 Nov;181(1):149–151. [PubMed] [Google Scholar]
- PRESCOTT D. M., BENDER M. A. Synthesis of RNA and protein during mitosis in mammalian tissue culture cells. Exp Cell Res. 1962 Mar;26:260–268. doi: 10.1016/0014-4827(62)90176-3. [DOI] [PubMed] [Google Scholar]
- ROTMAN B., SPIEGELMAN S. On the origin of the carbon in the induced synthesis beta-galactosidase in Escherichia coli. J Bacteriol. 1954 Oct;68(4):419–429. doi: 10.1128/jb.68.4.419-429.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SIMPSON M. V. The release of labeled amino acids from the proteins of rat liver slices. J Biol Chem. 1953 Mar;201(1):143–154. [PubMed] [Google Scholar]
- SIMPSON M. V., VELICK S. F. The synthesis of aldolase and glyceraldehyde-3-phosphate dehydrogenase in the rabbit. J Biol Chem. 1954 May;208(1):61–71. [PubMed] [Google Scholar]
- STEINBERG D., VAUGHAN M. Observations on intracellular protein catabolism studied in vitro. Arch Biochem Biophys. 1956 Nov;65(1):93–105. doi: 10.1016/0003-9861(56)90180-1. [DOI] [PubMed] [Google Scholar]
- SWICK R. W. Measurement of protein turnover in rat liver. J Biol Chem. 1958 Apr;231(2):751–764. [PubMed] [Google Scholar]
- Salb J. M., Marcus P. I. Translational inhibition in mitotic HeLa cells. Proc Natl Acad Sci U S A. 1965 Nov;54(5):1353–1358. doi: 10.1073/pnas.54.5.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schimke R. T., Sweeney E. W., Berlin C. M. Studies of the stability in vivo and in vitro of rat liver tryptophan pyrrolase. J Biol Chem. 1965 Dec;240(12):4609–4620. [PubMed] [Google Scholar]
- Schlessinger D., Ben-Hamida F. Turnover of protein in Escherichia coli starving for nitrogen. Biochim Biophys Acta. 1966 Apr 18;119(1):171–182. doi: 10.1016/0005-2787(66)90048-7. [DOI] [PubMed] [Google Scholar]
- URBA R. C. Protein breakdown in Bacillus cereus. Biochem J. 1959 Mar;71(3):513–518. doi: 10.1042/bj0710513. [DOI] [PMC free article] [PubMed] [Google Scholar]
