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Physical, immunological and kinetic properties of ribulose‐1,5‐bisphosphate carboxylase/oxygenase from fir (Abies alba) and spruce (Picea abies)
Author(s) -
Afif D.,
Gérant D.,
Cavalié G.,
Dizengremel P.
Publication year - 1993
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1993.tb01768.x
Subject(s) - picea abies , rubisco , abies alba , oxygenase , biology , botany , biochemistry , isoelectric focusing , ribulose , photosynthesis , chemistry , enzyme
Ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) from fir ( Abies alba Mill.) and spruce ( Picea abies [L.] Karst.) needles was purified to homogeneity. The enzyme was isolated from crude extracts through quantitative precipitation in 40‐55% and 40‐60% (NH 4 ) 2 SO 4 for fir and spruce. respectively, followed by linear sucrose gradient centrifugation. Using two dimensional gel electrophoresis, the isoelectric points were determined. For the large subunit (LSU) it was 6.7 for both species, and for the small subunit (SSU) it was 7.1 and 7.7 for fir and spruce, respectively. Very few differences in tryptic peptides and amino acid composition of Rubisco LSU were observed between fir and spruce. By contrast, marked differences characterized the same analyses for the Rubisco SSU of the two species. Moreover, substitution of residues was observed in the sequenced N‐terminal region when comparing fir and spruce SSU. The Ouchterlony technique showed no immu‐nochemical difference between Rubisco of fir and spruce when a rabbit antiserum to spinach Rubisco was used. The Eadie‐Hofstee plots of carboxylase activity indicated that the apparent K m (CO 2 ) were 31 and 36 μ M for the fir and spruce enzymes, respectively.