Chaperonin-Repairable Subtle Incompleteness of Protein Assembly Induced by a Substitution of Hydrogen with Deuterium : Effect of GroE on Deuterated Ribulose 1,5-Bisphosphate Carboxylase : PROTEINS, ENZYMES AND METABOLISM

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For investigating the effect of slight modification of proteins on their higher-ordered structure, and that of chaperonin on the functional assembly of proteins, we prepared partially deuterated ribulose 1,5-bisphosphate carboxylase (Rubisco) by cultivating Chlorella ellipsoidea in 100 mol% D_2O medium. Chlorella cells grown in the D_2O medium (D-Chlorella) contained almost the same amount of Rubisco (D-Rubisco) as the cells grown in H_2O medium (H-Chlorella) determined by Western blotting using Rubisco-specific antibody, whereas the activity of D-Rubisco determined by carbon fixation was only 28% of that of Rubisco from H-Chlorella (H-Rubisco). D-Rubisco, however, showed similar K_m and pH and temperature optima to those of H-Rubisco as well as similar antibody binding capability. The enzyme activity of D-Rubisco was recovered to 84% of that of H-Rubisco by the addition of GroE proteins (GroEL, chaperonin 60, and GroES, chaperonin 10), members of the chaperonin family produced by Escherichia coli. These data suggest that D-Rubisco has subtle incompleteness in terms of functional assembly, a situation that is correctable by chaperonin.
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