Structure and dynamics of a ribosome-bound nascent chain by NMR spectroscopy

被引:94
作者
Hsu, Shang-Te Danny
Fucini, Paola
Cabrita, Lisa D.
Launay, Helene
Dobson, Christopher M.
Christodoulou, John
机构
[1] Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England
[2] Max Planck Inst Mol Genet, D-14195 Berlin, Germany
[3] Univ Frankfurt, Inst Organ Chem & Chem Biol, D-60438 Frankfurt, Germany
基金
英国医学研究理事会; 英国惠康基金;
关键词
cotranslational folding;
D O I
10.1073/pnas.0704664104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 polypepticle chain is selectively C-13/N-15-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 (1g) domains. Analysis of these spectra reveals that the nascent chain adopts a structure in which a native-like N-tdrminal 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.
引用
收藏
页码:16516 / 16521
页数:6
相关论文
共 38 条
[1]   Function of trigger factor and DnaK in multidomain protein folding: Increase in yield at the expense of folding speed [J].
Agashe, VR ;
Guha, S ;
Chang, HC ;
Genevaux, P ;
Hayer-Hartl, M ;
Stemp, M ;
Georgopoulos, C ;
Hartl, FU ;
Barral, JM .
CELL, 2004, 117 (02) :199-209
[2]   ROTATIONAL DIFFUSION OF ESCHERICHIA-COLI RIBOSOMES .1. FREE 70 S-PARTICLE 50 S-PARTICLE AND 30 S-PARTICLE [J].
AMAND, B ;
POCHON, F ;
LAVALETTE, D .
BIOCHIMIE, 1977, 59 (10) :779-784
[3]   PRINCIPLES THAT GOVERN FOLDING OF PROTEIN CHAINS [J].
ANFINSEN, CB .
SCIENCE, 1973, 181 (4096) :223-230
[4]   Protein folding mechanisms: new methods and emerging ideas [J].
Brockwell, DJ ;
Smith, DA ;
Radford, SE .
CURRENT OPINION IN STRUCTURAL BIOLOGY, 2000, 10 (01) :16-25
[5]   Protein misfolding, functional amyloid, and human disease [J].
Chiti, Fabrizio ;
Dobson, Christopher M. .
ANNUAL REVIEW OF BIOCHEMISTRY, 2006, 75 :333-366
[6]   Heteronuclear NMR investigations of dynamic regions of intact Escherichia coli ribosomes [J].
Christodoulou, J ;
Larsson, G ;
Fucini, P ;
Connell, SR ;
Pertinhez, TA ;
Hanson, CL ;
Redfield, C ;
Nierhaus, KH ;
Robinson, CV ;
Schleucher, J ;
Dobson, CM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (30) :10949-10954
[7]   The present view of the mechanism of protein folding [J].
Daggett, V ;
Fersht, A .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2003, 4 (06) :497-502
[8]   NMRPIPE - A MULTIDIMENSIONAL SPECTRAL PROCESSING SYSTEM BASED ON UNIX PIPES [J].
DELAGLIO, F ;
GRZESIEK, S ;
VUISTER, GW ;
ZHU, G ;
PFEIFER, J ;
BAX, A .
JOURNAL OF BIOMOLECULAR NMR, 1995, 6 (03) :277-293
[9]   Understanding protein folding via free-energy surfaces from theory and experiment [J].
Dinner, AR ;
Sali, A ;
Smith, LJ ;
Dobson, CM ;
Karplus, M .
TRENDS IN BIOCHEMICAL SCIENCES, 2000, 25 (07) :331-339
[10]   The dynamic tunnel [J].
Etchells, SA ;
Hartl, FU .
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2004, 11 (05) :391-392