Enzymatic activity of the ribosome-bound nascent polypeptide.
Firefly luciferase was shown to be completely folded and thus enzymatically active immediately upon release from the ribosome [Kolb et al. (1994) EMBO J. 13, 3631-3637]. However, no luciferase activity was observed while full-length luciferase was attached to the ribosome as a peptidyl-tRNA, probably because the C-terminal portion of the enzyme is masked by the ribosome and/or ribosome-associated proteins. Here we have demonstrated that the ribosome-bound enzyme acquires the enzymatic activity when its C-terminus is extended by at least 26 additional amino acid residues. The results demonstrate that the acquisition of the final native conformation by a nascent protein does not need the release of the protein from the ribosome.
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, Russian Federation.
E V
EV
Makeyev
V A
VA
Kolb
A S
AS
Spirin
D001483:Base Sequence; D008156:Luciferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D014176:Protein Biosynthesis; D011487:Protein Conformation; D017510:Protein Folding; D011691:Puromycin / Q000494:pharmacology; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D014908:Triticum
D046911:Macromolecular Substances; D012346:RNA, Transfer, Amino Acyl; C019037:tRNA, peptidyl-; D011691:Puromycin; D008156:Luciferases