Folding of firefly luciferase during translation in a cell-free system. |
| |
Authors: | V A Kolb E V Makeyev A S Spirin |
| |
Affiliation: | Institute of Protein Research, Academy of Sciences of Russia, Moscow Region. |
| |
Abstract: | In vitro synthesis of firefly luciferase and its folding into an enzymatically active conformation were studied in a wheat germ cell-free translation system. A novel method is described by which the enzymatic activity of newly synthesized luciferase can be monitored continuously in the cell-free system while this protein is being translated from its mRNA. It is shown that ribosome-bound polypeptide chains have no detectable enzymatic activity, but that this activity appears within a few seconds after luciferase has been released from the ribosome. In contrast, the renaturation of denatured luciferase under identical conditions occurs with a half-time of 14 min. These results support the cotranslational folding hypothesis which states that the nascent peptides start to attain their native tertiary structure during protein synthesis on the ribosome. |
| |
Keywords: | |
|
|