PMID:1348957
| Citation |
Fisher, MT (1992) Promotion of the in vitro renaturation of dodecameric glutamine synthetase from Escherichia coli in the presence of GroEL (chaperonin-60) and ATP. Biochemistry 31:3955-63 |
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| Abstract |
The folding and assembly of dodecameric glutamine synthetase (GS) from Escherichia coli was examined in the absence and presence of the E. coli heat shock protein, GroEL (chaperonin-60). At nonphysiological temperatures (15-20 degrees C), unfolded GS spontaneously renatured to 80-90% of its original activity in the absence of GroEL. At near-physiological temperatures (37 degrees C), only 20-40% of the original activity returns. Under the latter solution conditions, GroEL and ATP enhance the extent of GS renaturation to 70-80% of the original activity at 37 degrees C. In the absence of ATP, GroEL arrests the renaturation of unfolded GS by forming a stable binary complex. The addition of ATP to this complex resulted in the release of GS subunits and formation of active dodecameric GS. The order of addition of ATP or unfolded GS to GroEL results in differences in the t1/2 values where half-maximal GS activity is attained. At a constant GS concentration, the formation of the GroEL.GS complex followed by ATP addition resulted in approximately a 2-fold increase in the observed t1/2 value compared to that observed when GroEL was preincubated with ATP before the GS renaturation reaction was initiated. These differences in renaturation rates may be related to binding affinity differences between the ATP-free and -bound GroEL conformer for unfolded or partially folded protein substrates [Badcoe, I. G., Smith, C. J., Wood, S., Halsall, D. J., Holbrook, J. J., Lund, P., & Clarke, A. R. (1991) Biochemistry 30, 9195-9200].(ABSTRACT TRUNCATED AT 250 WORDS) |
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| Keywords |
Adenosine Triphosphate/analogs & derivatives; Adenosine Triphosphate/pharmacology; Adenylyl Imidodiphosphate/pharmacology; Bacterial Proteins/pharmacology; Chaperonin 60; Electrophoresis, Agar Gel; Escherichia coli/enzymology; Glutamate-Ammonia Ligase/chemistry; Heat-Shock Proteins/pharmacology; Kinetics; Macromolecular Substances; Magnesium/pharmacology; Manganese/pharmacology; Protein Conformation; Protein Denaturation |
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