Summary
RuBisCo activase catalyzes the activation and maintains the activated state of the photosynthetic enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCo, EC 4.1.1.39). We employed antisera prepared against the RuBisCo holoenzyme purified from tobacco and RuBisCo activase isolated from spinach to determine the localization of these proteins in leaves of C3-type higher plants and green algae. In leaves ofVicia faba, both RuBisCo activase and RuBisCo are distributed throughout the chloroplast stroma. In contrast, RuBisCo activase and RuBisCo are predominantly localized to the pyrenoid in the green algaeChlamydomonas reinhardtii andColeochaete scutata. The co-immunolocalization of RuBisCo activase and RuBisCo to the pyrenoid in these two green algal species suggests that pyrenoid-localized RuBisCo is catalytically competent. We conclude that the pyrenoid functions as a unique metabolic compartment of the chloroplast in which the reactions of the photosynthetic carbon reduction pathway are initiated.
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Abbreviations
- RuBisCo:
-
ribulose 1,5-bisphosphate carboxylase/oxygenase
- RuBP:
-
ribulose 1,5-bisphosphate
- Pipes:
-
1,4-piperazinedi-ethanesulfonic acid
- BSA:
-
bovine serum albumin
- PBS:
-
phosphate-buffered saline
- TBS:
-
Tris-buffered saline
- IgG:
-
immunoglobulin G
- Ci :
-
inorganic carbon
References
Aizawa K, Miyachi S (1986) Carbonic anhydrase and CO2 concentrating mechanisms in microalgae and cyanobacteria. FEMS Microbiol Rev 39: 215–233
Badger MR, Kaplan A, Berry JA (1978) A mechanism for concentrating CO2 inChlamydomonas reinhardtii andAnabaena variabilis and its role in photosynthetic CO2 fixation. Carnegie Inst Yearbook 77: 251–261
Blank RJ, Trench RK (1988) Immunogold localization of ribulose-1,5-bisphosphate carboxylase-oxygenase inSymbiodinium kawagutii Trench et Blank, an endosymbiotic dinoflagellate. Endocyt Cell Res 5: 75–82
Ekman P, Lignell Å, Pedersén M (1989) Localization of ribulose-1,5-bisphosphate carboxylase/oxygenase inGracilaria secundata (Rhodophyta) and its role as a nitrogen storage pool. Bot Mar 32: 527–534
Graham LE (1985) The origin of the life cycle of land plants. Am Scientist 73: 178–186
Jordan DB, Chollet R (1983) Inhibition of ribulose bisphosphate carboxylase by substrate ribulose 1,5-bisphosphate. J Biol Chem 258: 13752–13758
—, Ogren WL (1983) Species variation in kinetic properties of ribulose 1,5-bisphosphate carboxylase/oxygenase. Arch Biochem Biophys 227: 425–433
Kajikawa H, Okada M, Ishikawa F, Okada Y, Nakayama K (1988) Immunochemical studies on ribulose 1,5-bisphosphate carboxylase/oxygenase in the chloroplasts of the marine algaBryopsis maxima. Plant Cell Physiol 29: 549–556
Kiss JZ, Vasconcelos AC, Triemer RE (1986) Paramylon synthesis and chloroplast structure associated with nutrient levels inEuglena (Euglenophyceae). J Phycol 22: 327–333
Lacoste-Royal G, Gibbs SP (1985)Ochromonas mitochondria contain a specific chloroplast protein. Proc Natl Acad Sci USA 82: 1456–1459
— — (1987) Immunocytochemical localization of ribulose-1,5-bisphosphate carboxylase in the pyrenoid and thylakoid region of the chloroplast ofChlamydomonas reinhardtii. Plant Physiol 83: 602–606
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Nature 227: 680–685
McKay RML, Gibbs SP (1989) Immunocytochemical localization of ribulose 1,5-bisphosphate carboxylase/oxygenase in light-limited and light-saturated cells ofChlorella pyrenoidosa. Protoplasma 149: 31–37
— — (1990) Phycoerythrin is absent from the pyrenoid ofPorphyridium cruentum: photosynthetic implications. Planta 180: 249–256
- - (1991) Composition and function of pyrenoids: cytochemical and immunocytochemical approaches. Can J Bot (in press)
Miziorko HM, Lorimer GH (1983) Ribulose-l,5-bisphosphate carboxylase-oxygenase. Ann Rev Biochem 52: 507–535
Mustardy L, Cunningham FX Jr, Gantt E (1990) Localization and quantitation of chloroplast enzymes and light-harvesting components using immunocytochemical methods. Plant Physiol 94: 334–340
Newman SM, Derocher J, Cattolico RA (1989) Analysis of chromophytic and rhodophytic ribulose-l,5-bisphosphate carboxylase indicates extensive structural and functional similarities among evolutionarily diverse algae. Plant Physiol 91: 939–946
Nisius A, Ruppel HG (1987) Immunocytological and chemical studies on the stromacentre-forming protein fromAvena plastids. Planta 171: 443–452
Osafune T, Sumida S, Ehara T, Hase E (1989) Three-dimensional distribution of ribulose-1,5-bisphosphate carboxylase/oxygenase in chloroplasts of actively photosynthesizing cell ofEuglena gracilis. J Electron Microsc 38: 399–402
—, Yokota A, Sumida S, Hase E (1990) Immunogold localization of ribulose-1,5-bisphosphate carboxylase with reference to pyrenoid morphology in chloroplasts of synchronizedEuglena gracilis cells. Plant Physiol 92: 802–808
Perchorowicz JT, Raynes DA, Jensen RG (1981) Light limitation of photosynthesis and activation of ribulose bisphosphate carboxylase in wheat seedlings. Proc Natl Acad Sci USA 78: 2985–2989
Portis ARTr (1990) Rubisco activase. Biochim Biophys Acta 1015: 15–28
—, Salvucci ME, Ogren WL (1986) Activation of ribulosebisphosphate carboxylase/oxygenase at physiological CO2 and ribulosebisphosphate concentrations by rubisco activase. Plant Physiol 82: 967–971
Reynolds ES (1963) The use of lead citrate at high pH as an electronopaque stain in electron microscopy. J Cell Biol 17: 208–212
Roesler KR, Ogren WL (1990) Primary structure ofChlamydomonas reinhardtii rubisco activase and evidence for a single polypeptide. Plant Physiol (in press)
Rother T, Acker G, Scheibe R (1988) Immunogold localization of chloroplast protein in spinach leaf mesophyll, epidermis, and guard cells. Bot Acta 101: 311–320
Salvucci ME (1989) Regulation of rubisco activity in vivo. Physiol Plant 77: 164–171
—, Portis AR Jr, Ogren WL (1985) A soluble chloroplast protein catalyzes ribulosebisphosphate carboxylase/oxygenase activation in vivo. Photosynth Res 7: 193–201
—, Werneke JM, Ogren WL, Portis AR Jr (1987) Purification and species distribution of rubisco activase. Plant Physiol 84: 930–936
Streusand VJ, Portis AR Jr (1987) Rubisco activase mediates ATP-dependent activation of ribulose bisphosphate carboxylase. Plant Physiol 85: 152–154
Vaughn KC (1987) Two immunological approaches to the detection of ribulose-1,5-bisphosphate carboxylase in guard cell chloroplasts. Plant Physiol 84: 188–196
—, Campbell EO, Hasegawa J, Owen HA, Renzaglia KS (1990) The pyrenoid is the site of ribulose 1,5-bisphosphate carboxylase/ oxygenase accumulation in the hornwort (Bryophyta:Anthocerotae) chloroplast. Protoplasma 156: 117–129
Vladimirova MG, Markelova AG, Semenenko VE (1982) Use of the cytoimmunofluorescent method to clarify localization of ribulosebisphosphate carboxylase in pyrenoids of unicellular algae. Fiziol Rast 29: 941–950 (English translation: Soviet Plant Physiol 30: 725–734)
Werneke JM, Chatfield JM, Ogren WL (1988 a) Catalysis of ribulosebisphosphate carboxylase/oxygenase activation by the product of a rubisco activase cDNA clone expressed inEscherichia coli. Plant Physiol 87: 917–920
—, Zielinski RE, Ogren WL (1988 b) Structure and expression of spinach leaf cDNA encoding ribulosebisphosphate carboxylase/ oxygenase activase. Proc Natl Acad Sci USA 85: 787–791
—, Chatfield JM, Ogren WL (1989) Alternative mRNA splicing generates the two ribulosebisphosphate carboxylase/oxygenase activase polypeptides in spinach andArabidopsis. Plant Cell 1: 815–825
Yokota A, Canvin DT (1986) Changes of ribulose bisphosphate carboxylase/oxygenase content, ribulose bisphosphate concentration, and photosynthetic activity during adaption of high-CO2 grown cells to low-CO2 conditions inChlorella pyrenoidosa. Plant Physiol 80: 341–345
Zielinski RE, Werneke JM, Jenkins ME (1989) Coordinate expression of rubisco activase and rubisco during barley leaf cell development. Plant Physiol 90: 516–521
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McKay, R.M.L., Gibbs, S.P. & Vaughn, K.C. RuBisCo activase is present in the pyrenoid of green algae. Protoplasma 162, 38–45 (1991). https://doi.org/10.1007/BF01403899
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DOI: https://doi.org/10.1007/BF01403899