Photosynthetica 1997, 33(12):211-218 | DOI: 10.1023/A:1006888506925

Proteolytic degradation of barley ribulose-1,5-bisphosphate carboxylase/oxygenase and recognition of the fragments by monoclonal antibodies

K. Demirevska-Kepova1, L. Simova-Stoilova1, S. Kyurkchiev2
1 Institute of Plant Physiology, Bulgarian Academy of Sciences, Sofia, Bulgaria
2 Institute of Biology and Immunology of Reproduction, Bulgarian Academy of Sciences, Sofia, Bulgaria

Limited proteolysis of barley ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) was effected by treatments with trypsin and Staphylococcus aureus strain V8 protease. Treatment of native RuBPCO with proteases resulted in the degradation of the large subunit (LS) of the enzyme. Trypsin cleaved three fragments from the LS but the S. aureus strain V8 protease cleaved only one. The small subunit (SS) was not affected. In the presence of 0.5 % sodium dodecyl sulfate, RuBPCO degraded into several fragments; some of them were fairly stable. Monoclonal antibodies (Mabs) against barley RuBPCO were applied in immunoblotting analysis to distinguish which of the fragments were recognized. All the Mabs recognized the fragments with molecular masses close to those of the LS. Differences among Mabs were observed in the fragments with low molecular mass.

Additional key words: Hordeum vulgare L.; limited proteolysis; monoclonal antibodies; RuBPCO

Prepublished online: January 1, 1998; Published: March 1, 1997  Show citation

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Demirevska-Kepova, K., Simova-Stoilova, L., & Kyurkchiev, S. (1997). Proteolytic degradation of barley ribulose-1,5-bisphosphate carboxylase/oxygenase and recognition of the fragments by monoclonal antibodies. Photosynthetica, 34(2), 211-218. doi: 10.1023/A:1006888506925
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    References

    1. Bottomley, W.: Characterization of chloroplast proteins by partial proteolytic digestion.-In: Edelman, M., Hallick, R.B., Chua, N.-H. (ed.): Methods in Chloroplast Molecular Biology. Pp. 1055-1061. Elsevier Biomedical Press, Amsterdam-New York-Oxford 1982.
    2. Bradford, M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding.-Anal. Biochem. 72: 248-254, 1976. Go to original source...
    3. Cleveland, D.W., Fischer, S.G., Kirschner, M.W., Laemmli, U.K.: Peptide mapping by limited proteolysis in SDS and analysis by gel electrophoresis.-J. biol. Chem. 252: 1102-1106, 1977. Go to original source...
    4. Demirevska-Kepova, K., Simova, L.: [Isolation and purification of ribulose 1,5-bisphosphate carboxylase/oxygenase from barley leaves.]-Fiziol. Rast. (Sofia) 15(3): 3-10, 1989. [In Bulg.]
    5. Demirevska-Kepova, K., Simova-Stoilova, L., Kyurkchiev, S., Juperlieva-Mateeva, B.: Epitope mapping of RuBPCO from barley leaves.-Compt. rend. Acad. Sci. Bulg. 49: 93-96, 1996.
    6. Desimone, M., Henke, A., Wagner, E.: Oxidative stress induces partial degradation of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase in isolated chloroplasts of barley.-Plant Physiol. 111: 789-796, 1996. Go to original source...
    7. Gutteridge, S., Millard, B.N., Parry, M.A.J.: Inactivation of ribulose-bisphosphate carboxylase by limited proteolysis: Loss of the catalytic activity without disruption of bisphosphate binding or carboxylation.-FEBS Lett. 196: 263-268, 1986. Go to original source...
    8. Hildbrand, M., Fischer, A., Feller, U.: Protein catabolism in bean leaf discs: accumulation of a soluble fragment of ribulose-1,5-bisphosphate carboxylase/oxygenase under oxygen deficiency.-J. exp. Bot. 45: 1197-1204, 1994. Go to original source...
    9. Houtz, R.L., Mulligan, R.M.: Protection of tryptic-sensitive sites in the large subunit of ribulosebisphosphate carboxylase/oxygenase by catalysis.-Plant Physiol. 96: 335-339, 1991. Go to original source...
    10. Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4.-Nature 227: 680-685, 1970. Go to original source...
    11. Mac, T., Kamei, C., Funaki, K., Miyadai, K., Makino, A., Ohira, K., Ojima, K.: Degradation of ribulose-1,5-bisphosphate carboxylase/oxygenase in the lysates of the chloroplasts isolated mechanically from wheat (Triticum aestivum L.) leaves.-Plant Cell Physiol. 30: 193-200, 1989. Go to original source...
    12. Mitsuhashi, W., Craft-Brandner, S.J., Feller, U.: Ribulose-1,5-bisphosphate carboxylase/oxygenase degradation in isolated pea chloroplasts incubated in the light or in the dark.-J. Plant Physiol. 139: 653-658, 1992. Go to original source...
    13. Mladjova, N., Demirevska-Kepova, K., Kehayov, J., Kyurkchiev, S.: Production and characterization of monoclonal antibodies to barley ribulose 1,5-bisphosphate carboxylase/oxygenase.-Plant Physiol. Biochem. 30: 523-529, 1992.
    14. Moreno, J., Peñarrubia, L., Garcia-Ferris, C.: The mechanism of redox regulation of ribulose 1,5-bisphosphate carboxylase/oxygenase turnover. A hypothesis.-Plant Physiol. Biochem. 33: 121-127, 1995.
    15. Mulligan, R.M., Houtz, R.L., Tolbert, N.E.: Reaction-intermediate analogue binding by ribulose bisphosphate carboxylase/oxygenase causes specific changes in proteolytic sensitivity: The N-terminal residue of the large subunit is acetylated proline.-Proc. nat. Acad. Sci. USA 85: 1513-1517, 1988. Go to original source...
    16. Otto, S., Feierabend, J.: Assay and comparative characterization of the protoolytic degradation of isolated small subunit and holoenzyme of ribulose-1,5-bisphosphate carboxylase/oxygenase in chloroplasts from rye leaves.-J. Plant Physiol. 144: 26-33, 1994. Go to original source...
    17. Sucyoshi, K., Kubo, Y., Yamagishi, K., Ogura, N., Ochiai, K., Fukushima, K., Ikeda, T., Nakagawa, H.: Monoclonal antibodies specific for NADH-ferricyanide reductase domain of spinach leaf nitrate reductase.-Plant Cell Physiol. 29: 975-980, 1988.
    18. Xu, Y., Van Huystee, R.B.: Identification of an antigenic determinant on anionic peanut peroxidase by monoclonal antibodies.-J. exp. Bot. 42: 935-945, 1991. Go to original source...

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