Factors permitting prolonged translation by isolated pea chloroplasts.

The following parameters were found to prolong the time-course of translation by isolated pea (Pisum sativum, cv Progress No. 9) chloroplasts: addition of other amino acids (an effect synergistic with sufficient free Mg(2+)), use of lower light intensities, and additions of inorganic phosphate and ATP. In a chloroplast system which includes these parameters, active translation usually extends to almost an hour. The total amount of leucine incorporated is routinely 60 to 100 nanomoles/milligram chlorophyll and often 200 nanomoles/milligram chlorophyll. Accurate estimation of the amount of amino acid incorporated depends on supplying the labeled amino acid at a concentration sufficient to overcome isotope dilution effects from endogenous pools. Approximately 39 thylakoid and 60 stroma polypeptides were visible on autoradiographs after labeling with [(35)S]methionine. Label in a few of the polypeptide bands was increased or decreased by specific changes in the reaction conditions. Due to the long period of activity and the large number of labeled products, this chloroplast system should be useful for future studies of chloroplast translation.

[1]  A. Jagendorf,et al.  Permeability of chloroplast envelopes to mg: effects on protein synthesis. , 1984, Plant physiology.

[2]  A. Jagendorf,et al.  ATP‐dependent proteolysis in pea chloroplasts , 1984 .

[3]  A. Goldberg,et al.  Newly synthesized proteins are degraded by an ATP‐stimulated proteolytic process in isolated pea chloroplasts , 1984 .

[4]  A. Jagendorf,et al.  A Mg2+ requirement for rapid ATP-driven protein synthesis by intact pea chloroplasts☆ , 1983 .

[5]  G. Fink,et al.  Positive regulation in the general amino acid control of Saccharomyces cerevisiae. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[6]  R. Huffaker,et al.  Evidence for the existence of peptide hydrolase activity associated with chloroplasts isolated from barley mesophyll protoplasts , 1983 .

[7]  Jan M. Anderson,et al.  Changes in composition and function of thylakoid membranes as a result of photosynthetic adaptation of chloroplasts from pea plants grown under different light conditions , 1983 .

[8]  B. Miflin,et al.  Intracellular localization of aspartate kinase and the enzymes of threonine and methionine biosynthesis in green leaves. , 1983, Plant physiology.

[9]  P. Milos,et al.  Light-dependent assembly of ribulose-1,5-bisphosphate carboxylase. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[10]  A. Jagendorf,et al.  High rates of protein synthesis by isolated chloroplasts. , 1982, Plant physiology.

[11]  M. Edelman,et al.  General occurrence and structural similarity of the rapidly synthesized, 32,000-dalton protein of the chloroplast membrane. , 1982, The Journal of biological chemistry.

[12]  A. Jagendorf,et al.  Light-induced increase in the number and activity of ribosomes bound to pea chloroplast thylakoids in vivo. , 1982, Plant physiology.

[13]  R. Ellis.  Chloroplast Proteins: Synthesis, Transport, and Assembly , 1981 .

[14]  R. Douce,et al.  Slow Passive Diffusion of Orthophosphate between Intact Isolated Chloroplasts and Suspending Medium. , 1981, Plant physiology.

[15]  A. Grossman,et al.  Energy-dependent uptake of cytoplasmically synthesized polypeptides by chloroplasts , 1980, Nature.

[16]  W. Mills.  Photosynthetic formation of the aspartate family of amino acids in isolated chloroplasts. , 1980, Plant physiology.

[17]  B. Miflin,et al.  Distribution of the Enzymes of Nitrogen Assimilation within the Pea Leaf Cell. , 1979, Plant physiology.

[18]  A. Rich,et al.  Membrane proteins synthesized but not processed by isolated maize chloroplasts , 1978, The Journal of cell biology.

[19]  W. Mills,et al.  Amino acid biosynthesis in isolated pea chloroplasts: metabolism of labeled aspartate and sulfate , 1978 .

[20]  J. Morgenthaler,et al.  Synthesis of soluble, thylakoid, and envelope membrane proteins by spinach chloroplasts purified from gradients. , 1976, Archives of biochemistry and biophysics.

[21]  D. Spencer,et al.  Protein synthesis in isolated spinach chloroplasts: comparison of light-driven and ATP-driven synthesis. , 1974, Archives of biochemistry and biophysics.

[22]  H. Kersten,et al.  Inhibitors of nucleic acid synthesis: biophysical and biochemical aspects. , 1974, Molecular biology, biochemistry, and biophysics.

[23]  R. Ellis,et al.  Protein synthesis in chloroplasts. I. Light-driven synthesis of the large subunit of fraction I protein by isolated pea chloroplasts. , 1973, Biochimica et biophysica acta.

[24]  R. Leech,et al.  Amino Acid Biosynthesis by Isolated Chloroplasts during Photosynthesis. , 1972, Plant physiology.

[25]  R. Mans,et al.  Measurement of the incorporation of radioactive amino acids into protein by a filter-paper disk method , 1961 .