Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation.
The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a +5 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of 35S-labelled ribulose bisphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose bisphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose bisphosphate carboxylase. For short periods of time (< 1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose bisphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photo-synthetic tissues.
Ferreira
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RM
Departamento de Botnica e Engenharia Biológica, Universidade Técnica de Lisboa, Portugal.
Franco
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Teixeira
A R
AR
D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D005635:Fructosediphosphates / Q000494:pharmacology; D005636:Fructosephosphates / Q000494:pharmacology; D015151:Immunoblotting; D007700:Kinetics; D008970:Molecular Weight; D010428:Pentosephosphates / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D010944:Plants / Q000201:enzymology*; D011487:Protein Conformation; D012273:Ribulose-Bisphosphate Carboxylase / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects*; D013402:Sugar Alcohols / Q000494:pharmacology; D014466:Ultraviolet Rays
D005635:Fructosediphosphates; D005636:Fructosephosphates; D010428:Pentosephosphates; D010455:Peptides; D013402:Sugar Alcohols; C032687:2-carboxyarabinitol 1,5-biphosphate; C027618:fructose-6-phosphate; D012273:Ribulose-Bisphosphate Carboxylase; C029063:fructose-1,6-diphosphate