GroEL reversibly binds to, and causes rapid inactivation of, human carbonic anhydrase II at high temperatures.
The initial yield of reactivation of GuHCl denatured human carbonic anhydrase II does not change with temperature between 3 and 35 degrees C. At temperatures above 35 degrees C, the enzymatic activity is not stable, but decreases over time. If the bacterial chaperonin GroEL is present during reactivation, the initial yield is lower compared to the spontaneous reaction at temperatures of 35-50 degrees C. However, unlike the spontaneous reactivation, the enzymatic activity with time in the presence of GroEL. In the presence of GroEL, native HCA II incubated at elevated temperatures will rapidly loose enzymatic activity to the same value as during reactivation at that particular temperature; most of the activity will recover if the temperature is lowered when GroEL is present. It is evident that there is an equilibrium between an inactive intermediate of HCA II, probably bound to GroEL, and active enzyme. Furthermore, proline isomerization is part of the rate-limiting step of refolding even in the presence of GroEL, and it is very noteworthy that prolyl isomerase will influence the refolding of HCA II in the presence of GroEL.
Persson
M
M
IFM/Dept. of Chemistry, Linköping University, Sweden.
Carlsson
U
U
Bergenhem
N C
NC
D000591:Amino Acid Isomerases / Q000378:metabolism; D002256:Carbonic Anhydrases / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D018834:Chaperonin 60 / Q000378:metabolism*; D004789:Enzyme Activation; D004926:Escherichia coli; D019791:Guanidine; D006146:Guanidines / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D019696:Peptidylprolyl Isomerase; D011485:Protein Binding; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism; D013696:Temperature
D002352:Carrier Proteins; D018834:Chaperonin 60; D006146:Guanidines; D011994:Recombinant Proteins; D002256:Carbonic Anhydrases; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase; D019791:Guanidine