Summary
Isoelectric focusing of subunits of ribulose 1,5-bisphosphate carboxylase oxygenase of Medicago, Spinacia and Nicotiana were investigated, using a rapid isolation technique, without S-carboxymethylation. RuBPC-ase and its subunits were isolated by gel electrophoresis. Isoelectric focusing of RuBPC-ase of M. sativa and M. falcata showed that this enzyme consists of one large subunit (LSU) polypeptide and two or three small subunits (SSU), depending on the genotype. The pl of the LSU's was identical, but the pl of SSU's of the two genotypes was different. Amino acid composition and tryptic peptide maps further supported the concept of a conserved nature of LSU and heterogeneity of SSU polypeptides in Medicago. It was also found that S. oleracea, N. tabacum, N. glutinosa and N. excelsior have a single LSU polypeptide, but they differ in respect of pl values. The SSU polypeptides appeared to be variable. S-carboxymethylation affected the number as well as the pl values of LSU and SSU polypeptides. It is suggested that one LSU polypeptide is probably the general rule in higher plants, rather than the three LSU polypeptides demonstrated by Chen et al. (1977) and Wildman (1979).
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References
Amiri I, Salnikow J, Vater J (1984) Amino acid sequence of the large subunit of d ribulose bisphosphate carboxylase oxygenase EC — 4.1.1.39 from Nicotiana tabacum. Biochim Biophys Acta 784:116–123
Bedbrook JR, Coen DM, Beaton AR, Bogorad L, Rich A (1979) Location of single gene for the large subunit of ribulose bisphosphate carboxylase on the maize chloroplast chromosome. J Biol Chem 254:905–910
Bedbrook JR, Smith SM, Ellis J (1980) Molecular cloning and sequencing of cDNA encoding the precursor to the small subunit of chloroplast ribulose-1,5-bisphosphate carboxylase. Nature 287:692–697
Berry-Lowe SL, McKnight TD, Shah DM, Meagher RB (1982) The nucleotide sequence, expression and evolution of one member of a multigene family encoding the small subunit of ribulose-1,5-bisphosphate carboxylase in soybean. J Mol Appl Genet 1:483–498
Bovenberg WA, Koes RE, Kool AJ, Nijkamp HJJ (1984) Physical mapping nucleotide sequencing and expression in Escherichia coli mini cells of the gene for the large subunit of ribulose bisphosphate carboxylase from Petunia hybrida. Curr Genet 8:231–241
Byers M (1971) Amino acid composition of some leaf protein preparations. In: Pirie NW (ed) Leaf protein: its agronomy, preparation, quality and use. IBP Blackwell, Oxford Edinburgh, pp 95–114
Cammaerts D, Jacobs M (1981) Genetic control of small subunit polypeptides of Fraction 1 protein in the genus Lycopersicum. Plant Sci Lett 22:147–154
Chan PH, Wildman SG (1972) Chloroplast DNA codes for the primary structure of the large subunit of Fraction 1 protein. Biochim Biophys Acta 277:677–680
Chen K, Johal S, Wildman SG (1977) Phenotypic markers for chloroplast DNA genes in higher plants and their use in biochemical genetics. In: Bogorad L, Weil JH (eds) Nucleic acid and protein synthesis in plants. Plenum Press, New York London, pp 183–194
Daday HV, Whitecross MI (1983) Isoelectric focusing of ribulose bisphosphate carboxylase from lucerne and spinach. Sci Tools 30:11–14
Davis BJ (1964) Disc electrophoresis. 11. Method and application to human serum proteins. Ann NY Acad Sci 121:404–427
Dron M, Rahire M, Rochaix JD (1982) Sequence of the chloroplast DNA region of Chlamydomonas — Reinhardii containing the gene of the large subunit of ribulose bisphosphate carboxylase and parts of its flanking genes. J Mol Biol 162:775–794
Dunsmuir P, Smith S, Bedbrook J (1983) A number of different nuclear genes for the small subunit of RuBPCase are transcribed in petunia. Nucleic Acids Res 11:4177–4183
Erion JL, Tarnowski J, Peacock S, Caldwell P, Redfield B, Brot N, Weissbach H (1983) Synthesis of the large subunit of ribulose-1,5-bisphosphate carboxylase in an in vitro partially defined E. coli system. Plant Mol Biol 2:279–290
Gray JC, Kung SD, Wildman SG (1978) Polypeptide chains of the large and small subunits of Fraction 1 protein from tobacco. Arch Biochem Biophys 185:272–281
Haslett BG, Yarwood A, Evans IM, Boulter D (1976) Studies on the small subunit of Fraction 1 protein from Pisum sativum L. and Vicia faba L. Biochim Biophys Acta 420:122–132
Kawashima N, Wildman SG (1970) A model of the subunit structure of Fraction 1 protein. Biochem Biophys Res Commun 41:1463–1468
Kawashima N, Wildman SG (1972) Studies on Fraction 1 protein. 4. Mode of inheritance of primary structure in relation to whether choroplast or nuclear DNA contains the code for chloroplast protein Biochim Biophys Acta 262:42–49
Kung SD (1984) Genetic analysis of tobacco RuBPCase. CRC Crit Rev Plant Sci 1:227–267
Martin PG, Dowd JM (1984) The study of plant phylogeny using amino-acid sequences of ribulose-1,5-bisphosphate carboxylase. 3. Addition of Malvaceae and Ranunculaceae to phylogenetic tree. Aust J Bot 32:283–290
McIntosh L, Poulsen C, Bogorad L (1980) Chloroplast gene sequence for the large subunit of ribulose bisphosphate carboxylase of maize. Nature 288:556–560
Mueller KD, Salnikow J, Vater J (1983) Amino acid sequence of small subunit of d ribulose bisphosphate carboxylase EC-4. 1.1.39 oxygenase from Nicotiana tabacum. Biochim Biophys Acta 742:78–83
Noguchi H, Maekawa T, Fujimoto S, Stake I, Sakakibara M (1978) Physicochemical studies on Fraction 1 protein from alfalfa. Agric Biol Chem 42:1553–1558
O'Connell PBH, Brady CJ (1981) Multiple forms of the large subunit of wheat ribulose bisphosphate carboxylase generated by excess iodoacetamide. Biochim Biophys Acta 670:355–361
Poulsen C (1981) Comments on the structure and function of the large subunit of the enzyme ribulose bisphosphate carboxylase oxygenase EC 4.1.1.39. Carlsberg Res Commun 46:259–278
Smith SM, Speirs J, Dunsmuir P, Bedbrook J (in preparation) An invariant hexadecapeptide in the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from four species of higher plant
Wettstein D von, Poulsen C, Holder AA (1978) Ribulose-1,5-bisphosphate carboxylase as a nuclear and chloroplast marker. Theor Appl Genet 53:193–197
Wildman SG (1979) Aspects of Fraction 1 protein evolution. Arch Biochem Biophys 196:598–610
Zurawski G, Perrot B, Bottomley W, Whitfeld PR (1981) The structure of the gene for the large subunit of ribulose 1,5- bisphosphate carboxylase from spinach chloroplast DNA. Nucleic Acids Res 9:3251–3269
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Communicated by H. F. Linskens
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Daday, H.V., Whitecross, M.I. & Shaw, D.C. One large subunit of ribulose 1,5-bisphosphate carboxylase oxygenase in Medicago, Spinacia and Nicotiana . Theoret. Appl. Genetics 71, 708–715 (1986). https://doi.org/10.1007/BF00263268
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DOI: https://doi.org/10.1007/BF00263268