Supplemental Data

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• Supplemental Figure 1 - (06-7958SupplementalFig1.TIF; 30 KB) Addition of GroES and ADP to GroEL-RUBISCO binary complex causes protection to RUBISCO against proteinase K digestion. ³⁵ S-labelled RUBISCO, denatured in 2 M GdnHCl, was diluted by 90-folds with the refolding buffer containing GroEL such that the final concentration of RUBISCO and GroEL was 1μM in the solution. To form GroEL-RUBISCO binary complex, the diluted solution was incubated for 10 min at 25° C. GroEL-RUBISCO binary sample was then divided into two parts. ADP (5 mM) along with GroES (2 μM) was added to the second sample and nothing was added in the first sample. The samples were subsequently treated with 5 μg/ml proteinase K (which was previously optimized) at 25° C for 10 min and the digestion reactions were quenched with the addition of PMSF to a final concentration of 1 mM. The digested mixtures were run on a 10% SDS-PAGE gel and processed further. Appearance of RUBISCO band in a proteinase K treated sample is an indication for the resistance of the protein from proteolytic digestion. GroEL-RUBISCO binary complex (Lane 1), GroEL-RUBISCO binary complex with proteinase K (Lane 2), GroEL-RUBISCO binary complex in presence of GroES and ADP (in absence and presence of proteinase K, Lanes 3 and 4).