Identification of the Large Subunit of Ribulose 1,5-Bisphosphate Carboxylase/Oxygenase as a Substrate for Transglutaminase in Medicago sativa L. (Alfalfa)

Autor:	Megan R. Bruce-Carver, Donna Louie, Andrea P. Gonzales, Abdi Dharma, Stephen A. Margosiak, Glenn D. Kuehn
Rok vydání:	1990
Předmět:	Gel electrophoresis chemistry.chemical_classification Oxygenase Ribulose 1 5-bisphosphate biology Physiology Protein subunit Ribulose RuBisCO food and beverages Plant Science Pyruvate carboxylase chemistry.chemical_compound Enzyme chemistry Biochemistry Genetics biology.protein
Zdroj:	Plant Physiology. 92:88-96
ISSN:	1532-2548 0032-0889
DOI:	10.1104/pp.92.1.88
Popis:	Extracts prepared from floral meristematic tissue of alfalfa (Medicago sativa L.) were investigated for expression of the enzyme transglutaminase in order to identify the major protein substrate for transglutaminase-directed modifications among plant proteins. The large polymorphic subunits of ribulose 1,5-bisphosphate carboxylase/oxygenase in alfalfa, with molecular weights of 52,700 and 57,600, are major substrates for transglutaminase in these extracts. This was established by: (a) covalent conjugation of monodansylcadaverine to the large subunit followed by fluorescent detection in SDS-polyacrylamide gels; (b) covalent conjugation of [14C]putrescine to the large subunit with detection by autoradiography; (c) covalent conjugation of monodansylcadaverine to the large subunit and demonstration of immunocross-reactivity on nitrocellulose transblot of the modified large subunit with antibody prepared in rabbits against dansylated-ovalbumin; (d) demonstration of a direct dependence of the rate of transglutaminase-mediated, [14C]putrescine incorporation upon the concentration of ribulose, 1,5-bisphosphate carboxylase/oxygenase from alfalfa or spinach; and (e) presumptive evidence from size exclusion chromatography that transglutaminase may cofractionate with native molecules of ribulose 1,5-bisphosphate carboxylase/oxygenase in crude extracts. Analysis of the primary structure of plant large subunit has revealed numerous potential glutaminyl and lysyl sites for transglutaminase-directed modifications of ribulose 1,5-bisphosphate carboxylase/oxygenase.
Databáze:	OpenAIRE
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