Abstract
Ribulose bisphosphate (RuP2) carboxylase from the marme cyanobacterium, Synechococcus sp., comprised both large (57,000 dalton) and small (12,000 dalton) subunits. The undissociated, purified enzyme was considerably smaller than the spinach enzyme when compared by pore-gradient electrophoresis, gel filtration and density-gradient centrifugation. This suggested that the cyanobacterial enzyme might have a hexameric (L6S6) subunit structure, unlike the enzymes from spinach and many other organisms which are octamers (L8S8). However, the molecular weight of the Synechococcus enzyme was measured by equilibrium sedimentation and found to be 530,000, which is within the range observed for L8S8-type enzymes. Furthermore, electron microscopic studies of negatively stained preparations of both the native enzyme, and a preparation depleted of 87% of its small subunits by repeated mild-acid precipitation, revealed four-fold symmetry characteristic of an octameric, cubical structure. Synechococcus RuP2 carboxylase therefore must be an L8S8 octamer and its anomalous pore-penetration behaviour may be due to an asymmetric shape. Some support for the latter possibility was provided by electron miscoscopic observations of two different types of images which may be different views of the molecule in two planes.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- RuP2 :
-
d-ribulose-1,5-bisphosphate
References
Ackers GK (1967) A new calibration procedure for gel filtration columns. J Biol Chem 242:3237–3238
Andrews TJ, Abel KM (1981) Kinetics and subunit interactions of ribulose bisphosphate carboxylase-oxygenase from the cyanobacterium, Synechococcus sp. J Biol Chem 256:8445–8451
Axelsson I (1978) Characterization of proteins and other macromolecules by agarose gel chromatography. J Chromatog 152:21–32
Bowien B, Mayer F, Codd GA, Schlegel HG (1976) Purification, some properties and quaternary structure of the d-ribulose 1,5-diphosphate carboxylase of Alcaligenes eutrophus. Arch Microbiol 110:157–166
Bradford MM (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principal of protein-dye binding. Analyt Biochem 72:248–254
Cohn EJ, Edsall JT (1943) Proteins, amino acids and peptides. Reinhold Publishing Corp, New York, pp 370–381
Edelhoch H (1967) Spectroscopic determination of tryptophan and tyrosine in proteins. Biochem 6:1948–1954
Felgenhauer K (1974) Evaluation of molecular size by gel electrophoretic techniques. Hoppe-Seyler's Z Physiol Chem 355:1281–1290
Lawlis VB, Gordon GLR, McFadden BA (1979) Ribulose 1,5-bisphosphate carboxylase/oxygenase from Pseudomonas oxalaticus. J Bacteriol 139:287–298
Lorimer GH, Andrews TJ (1981) The C2 chemo- and photorespiratory carbon oxidation cycle. In: MD Hatch, NK Boardman (eds) The biochemistry of plants, Vol. 8. Academic Press, New York, pp 329–374
Lorimer GH, Badger MR, Andrews TJ (1976) The activation of ribulose-1,5-bisphosphate carboxylase by carbon dioxide and magnesium ions. Equilibrium, kinetics, a suggested mechanism, and physiological implications. Biochem 15:529–536
Markham R, Frey S, Hills GJ (1963) Methods for the enhancement of image detail and accentuation of structure in electron microscopy. Virology 20:88–102
Martin RG, Ames BN (1961) A method for determining the sedimentation behaviour of enzymes: Application to protein mixtures. J Biol Chem 236:1372–1379
McFadden BA (1980) A perspective of ribulose biphosphate carboxylase/oxygenase, the key catalyst in photosynthesis and photorespiration. Acc Chem Res 13:394–399
Moore S (1963) On the determination of cystine as cysteic acid. J Biol Chem 238:235–237
Nozaki Y, Schecter NM, Reynolds JA, Tanford C (1976) Use of gel chromatography for the determination of the Stokes radii of proteins in the presence and absence of detergents. A reexamination. Biochem 15:3884–3890
Purohit K, McFadden BA, Cohen AL (1976) Purification, quaternary structure, compositon, and properties of d-ribulose-1,5-bisphosphate carboxylase from Thiobacillus intermedius. J Bacteriol 127:505–515
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61
Scott K, Cannell GR, Zerner B (1975) A computer program for processing data from amino acid analysis and for calculation of molecular weights from these data. Analyt Biochem 69:474–484
Siegel LM, Monty KJ (1966) Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta 112:346–362
Steer MW, Gunning BES, Graham TA, Carr DJ (1968) Isolation, properties and structure of fraction 1 protein from Avena sativa L. Planta 79:254–267
Taylor S, Dalton H, Dow C (1980) Purification and initial characterisation of ribulose 1,5-bisphosphate carboxylase from Methylococcus capsulatus (Bath.) FEMS Microbiol Lett 8:157–160
Taylor SC, Dow CS (1980) Ribulose-1,5-bisphosphate carboxylase from Rhodomicrobium vanniellii. J Gen Microbiol 116:81–87
Van Holde KE, Baldwin RL (1958) Rapid attainment of sedimentation equilibrium. J Phys Chem 62:734–743
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Andrews, T.J., Abel, K.M., Menzel, D. et al. Molecular weight and quaternary structure of ribulose bisphosphate carboxylase from the cyanobacterium, Synechococcus sp.. Arch. Microbiol. 130, 344–348 (1981). https://doi.org/10.1007/BF00414597
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00414597