Identification of the large subunit of Ribulose 1,5-bisphosphate carboxylase/oxygenase as a substrate for transglutaminase in Medicageo sativa L. (alfalfa)
- New Mexico State Univ., Las Cruces (USA)
Extract 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 ({sup 14}C)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, ({sup 14}C)putresciene 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.
- OSTI ID:
- 7100913
- Journal Information:
- Plant Physiology; (USA), Journal Name: Plant Physiology; (USA) Vol. 92:1; ISSN PLPHA; ISSN 0032-0889
- Country of Publication:
- United States
- Language:
- English
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59 BASIC BIOLOGICAL SCIENCES
ALFALFA
AUTORADIOGRAPHY
CARBON 14 COMPOUNDS
CARBON-CARBON LYASES
CARBOXY-LYASES
ELECTROPHORESIS
ENZYME ACTIVITY
ENZYMES
GENES
LABELLED COMPOUNDS
LEGUMINOSAE
LYASES
MAGNOLIOPHYTA
MAGNOLIOPSIDA
OXIDOREDUCTASES
OXYGENASES
PLANTS
POST-TRANSLATION MODIFICATION
RIBULOSE DIPHOSPHATE CARBOXYLASE
TRANSFERASES