Abstract
For purifying carboxysomes of Thiobacillus neapolitanus an isolation procedure was developed which resulted in carboxysomes free from whole cells, protoplasts and cell fragments. These purified carboxysomes are composed of 8 proteins and at the most of 13 polypeptides. The two most abundant proteins which make up more than 60% of the carboxysomes, are ribulose-1,5-bisphosphate carboxylase and a glycoprotein with a molecular weight of 54,000. The shell of the carboxysomes consists of four glycoproteins, one also with a molecular weight of 54,000. The other proteins are present in minor quantities. Ribulose-1,5-bisphosphate carboxylase is the only enzyme which could be detected in the carboxysomes and 3-phosphoglycerate was the only product formed during incubation with ribulose-1,5-diphosphate and bicarbonate. The supernatant of a broken and centrifuged carboxysome suspension contained the large subunit of ribulose-1,5-bisphosphate carboxylase. The small subunit of ribulose-1,5-bisphosphate carboxylase was found in the pellet together with the shell proteins which indicates that the small subunit of ribulose-1,5-bisphosphate carboxylase is connected to the shell.
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Abbreviations
- RuBisCO:
-
ribulose-1,5-bisphosphate carboxylase
- PMSF:
-
phenylmethylsulfonyl fluoride
- PAA:
-
gelectrophoresis, polyacrylamide gelelectrophoresis
- SDS:
-
sodium dodecyl sulphate
- CIE:
-
crossed immunoelectrophoresis
- IEF:
-
isoelectric focusing
References
Amesz GFL, Nikaido K (1976) Two dimensional gel electrophoresis of membrane proteins. Biochemistry 15:616–623
Beudeker RF, Codd GA, Kuenen JG (1981) Quantification and intracellular distribution of ribulose-1,5-bisphosphate carboxylase in Thiobacillus neapolitanus, as related to possible functions of carboxysomes. Arch Microbiol 129:361–367
Beudeker RF, Kuenen JG (1981) Carboxysomes: “Calvinosomes”?. FEBS Lett 131:269–274
Biedermann M, Westphal K (1979) Chemical composition of NB1 particles from Nitrobacter agilis. Arch Microbiol 121:187–191
Broekhuyse RM (1969) Quantitative two-dimensional thin layer chromatography of blood phospholipids. Clin Chim Acta 23: 457–461
Brown HM, Bowman LH, Chollet R (1981) An improved purification protocol for ribulose-1,5-bisphosphate carboxylase from Chromatium vinosum. FEMS Microbiol Lett 12:105–109
Cannon GC, Shively JM (1983) Characterization of a homogeneous preparation of carboxysomes from Thiobacillus neapolitanus. Arch Microbiol 134:52–59
Codd GA, Stewart WDP (1976) Polyhedral bodies and ribulose 1,5-diphosphate carboxylase of the blue-green alga Anabaena cylindrica. Planta 130:323–326
Codd GA, Marsden WJN (1984) The carboxysomes (polyhedral bodies) of autotrophic prokaryotes. Biol Rev 59:389–422
Elferink MGL, Hellingwerf KJ, Michels PAM, Seyen HG, Konings WN (1979) Immunochemical analysis of membrane vesicles and chromophores of Rhodopseudomonas sphaeroides by crossed immunoelectrophoresis. FEBS Lett 107:300–307
Holthuijzen YA, Breemen JFL van, Konings WN, Bruggen EFJ van (1986) Electron microscopic studies of the carboxysomes of Thiobacillus neapolitanus Arch Microbiol (in press)
Kuenen JG, Veldkamp H (1973) Effects of organic compounds on growth of chemostat cultures of Thiomicrospira pelophila, Thiobacillus thioparus and Thiobacillus neapolitanus. Arch Microbiol 94:173–190
Lanaras T, Codd GA (1981) Ribulose-1,5-bisphosphate carboxylase and polyhedral bodies of Chlorogloeopsis fritschii. Planta 153:279–285
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond) 227: 680–685
Laemmli UK, Favre K (1973) Maturation of the head of bacteriophage T4. I. DNA packing events. J Mol Biol 80:575–599
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. Biochemistry 15:616–623
Plas J van der, Hellingwerf KJ, Seyen HG, Guest JR, Weiner JH, Konings WN (1983) Identification and localization of enzymes of the fumarate reductase and nitrate respiration systems of Escherichia coli by crossed immunoelectrophoresis. J Bacteriol 153:1027–1037
Purohit K, McFadden BA, Shaykh MM (1976) d-Ribulose-1,5-bisphosphate carboxylase and polyhedral bodies in Thiobacillus intermedius. J Bacteriol 127:516–522
Rouser G, Fleischer S, Yamamoto A (1970) Two-dimensional thin layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots. Lipids 5:494–496
Segrest JP, Jackson RL (1972) Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulphate. Meth Enzymol 28B:54–63
Segrest JP, Jackson RL, Andrews EP, Marchesi VT (1971) Human erythrocyte membrane glycoprotein: a re-evaluation of the molecular weight as determined by SDS polyacrylamide gel electrophoresis. Biochem Biophys Res Commun 44:390–395
Shively JM (1974) Inclusion bodies of prokaryotes. Annu Rev Microbiol 28:167–188
Shively JM, Ball FL, Kline BW (1973) Electron microscopy of the carboxysomes (polyhedral bodies) of Thiobacillus neapolitanus. J Bacteriol 116:1405–1411
Smyth CJ, Siegel J, Salton MRJ, Owen P (1978) Immunochemical analysis of inner and outer membranes of Escherichia coli by crossed immunoelectrophoresis. J Bacteriol 133:306–319
Snead RM, Shively JM (1978) d-Ribulose-1,5-bisphosphate carboxylase from Thiobacillus neapolitanus. Curr Microbiol 1: 309–314
Stewart WDP, Codd GA (1975) Polyhedral bodies (carboxysomes) of nitrogen-fixing blue-green algae. British Phycol J 10:273–278
Thompson J, Thomas TD (1977) Phosphoenolpyruvate and 2-phosphoglycerate: endogenous energy source(s) for sugar accumulation by starved cells of Streptococcus lactis. J Bacteriol 1301:583–595
Vishniac W, Santer M (1957) The thiobacilli. Bacteriol Rev 21: 195–213
Wagner H, Hörhammer L, Wolff P (1961) Dünnschichtchromatographie von Phosphatiden und Glycolipiden. Biochem Z 334:175–184
Wary W, Boulikas T, Wray VP, Hancock R (1981) Silver staining of proteins in polyacrylamide gels. Anal Biochem 118:197–203
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Holthuijzen, Y.A., van Breemen, J.F.L., Kuenen, J.G. et al. Protein composition of the carboxysomes of Thiobacillus neapolitanus . Arch. Microbiol. 144, 398–404 (1986). https://doi.org/10.1007/BF00409891
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DOI: https://doi.org/10.1007/BF00409891