Structure and dynamics of a ribosome-bound nascent chain by NMR spectroscopy.
Protein folding in living cells is inherently coupled to protein synthesis and chain elongation. There is considerable evidence that some nascent chains fold into their native structures in a cotranslational manner before release from the ribosome, but, despite its importance, a detailed description of such a process at the atomic level remains elusive. We show here at a residue-specific level that a nascent protein chain can reach its native tertiary structure on the ribosome. By generating translation-arrested ribosomes in which the newly synthesized polypeptide chain is selectively (13)C/(15)N-labeled, we observe, using ultrafast NMR techniques, a large number of resonances of a ribosome-bound nascent chain complex corresponding to a pair of C-terminally truncated immunoglobulin (Ig) domains. Analysis of these spectra reveals that the nascent chain adopts a structure in which a native-like N-terminal Ig domain is tethered to the ribosome by a largely unfolded and highly flexible C-terminal domain. Selective broadening of resonances for a group of residues that are colocalized in the structure demonstrates that there are specific but transient interactions between the ribosome and the N-terminal region of the folded Ig domain. These findings represent a step toward a detailed structural understanding of the cellular processes of cotranslational folding.
Hsu
Shang-Te Danny
ST
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
Fucini
Paola
P
Cabrita
Lisa D
LD
Launay
Hélène
H
Dobson
Christopher M
CM
Christodoulou
John
J
D002247:Carbon Isotopes / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry / Q000235:genetics; D007553:Isotope Labeling; D009587:Nitrogen Isotopes / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry; D012270:Ribosomes / Q000737:chemistry*
D002247:Carbon Isotopes; D007136:Immunoglobulins; D009587:Nitrogen Isotopes; D011506:Proteins; D012333:RNA, Messenger; D004247:DNA