Direct observation of the three-state folding of a single protein molecule

被引:486
作者
Cecconi, C
Shank, EA
Bustamante, C
Marqusee, S
机构
[1] Univ Calif Berkeley, Dept Mol & Cell Biol, Berkeley, CA 94720 USA
[2] Univ Calif Berkeley, Inst Quantitat Biol, Berkeley, CA 94720 USA
[3] Univ Calif Berkeley, Dept Phys, Berkeley, CA 94720 USA
[4] Univ Calif Berkeley, Howard Hughes Med Inst, Berkeley, CA 94720 USA
关键词
D O I
10.1126/science.1116702
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
We used force-measuring optical tweezers to induce complete mechanical unfolding and refolding of individual Escherichia-coli ribonuclease H (RNase H) molecules. The protein unfolds in a two-state manner and refolds through an intermediate that correlates with the transient molten globule-like intermediate observed in bulk studies. This intermediate displays unusual mechanical compliance and unfolds at substantially lower forces than the native state. In a narrow range of forces, the molecule hops between the unfolded and intermediate states in real time. Occasionally, hopping was observed to stop as the molecule crossed the folding barrier directly from the intermediate, demonstrating that the intermediate is on-pathway. These studies allow us to map the energy landscape of RNase H.
引用
收藏
页码:2057 / 2060
页数:4
相关论文
共 29 条
  • [1] Is protein folding hierarchic? II. Folding intermediates and transition states
    Baldwin, RL
    Rose, GD
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1999, 24 (02) : 77 - 83
  • [2] Can non-mechanical proteins withstand force? Stretching barnase by atomic force microscopy and molecular dynamics simulation
    Best, RB
    Li, B
    Steward, A
    Daggett, V
    Clarke, J
    [J]. BIOPHYSICAL JOURNAL, 2001, 81 (04) : 2344 - 2356
  • [3] Pulling geometry defines the mechanical resistance of a β-sheet protein
    Brockwell, DJ
    Paci, E
    Zinober, RC
    Beddard, GS
    Olmsted, PD
    Smith, DA
    Perham, RN
    Radford, SE
    [J]. NATURE STRUCTURAL BIOLOGY, 2003, 10 (09) : 731 - 737
  • [4] Mechanical processes in biochemistry
    Bustamante, C
    Chemla, YR
    Forde, NR
    Izhaky, D
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 2004, 73 : 705 - 748
  • [5] The mechanical stability of ubiquitin is linkage dependent
    Carrion-Vazquez, M
    Li, HB
    Lu, H
    Marszalek, PE
    Oberhauser, AF
    Fernandez, JM
    [J]. NATURE STRUCTURAL BIOLOGY, 2003, 10 (09) : 738 - 743
  • [6] Detection of rare partially folded molecules in equilibrium with the native conformation of RNaseH
    Chamberlain, AK
    Handel, TM
    Marqusee, S
    [J]. NATURE STRUCTURAL BIOLOGY, 1996, 3 (09): : 782 - 787
  • [7] Chan HS, 1998, PROTEINS, V30, P2, DOI 10.1002/(SICI)1097-0134(19980101)30:1<2::AID-PROT2>3.0.CO
  • [8] 2-R
  • [9] EQUILIBRIUM UNFOLDING OF ESCHERICHIA-COLI RIBONUCLEASE-H - CHARACTERIZATION OF A PARTIALLY FOLDED STATE
    DABORA, JM
    MARQUSEE, S
    [J]. PROTEIN SCIENCE, 1994, 3 (09) : 1401 - 1408
  • [10] Force-clamp spectroscopy monitors the folding trajectory of a single protein
    Fernandez, JM
    Li, HB
    [J]. SCIENCE, 2004, 303 (5664) : 1674 - 1678