DOI: 10.1073/pnas.97.26.14241. Fluorescence quenching: A tool for single-molecule protein-folding study

Fluorescence quenching: A tool for single-molecule protein-folding study

10.1073/pnas.97.26.14241

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

By using titin as a model system, we have demonstrated that fluorescence quenching can be used to study protein folding at the single molecule level. The unfolded titin molecules with multiple dye molecules attached are able to fold to the native state. In the native folded state, the fluorescence from dye molecules is quenched due to the close proximity between the dye molecules. Unfolding of the titin leads to a dramatic increase in the fluorescence intensity. Such a change makes the folded and unfolded states of a single titin molecule clearly distinguishable and allows us to measure the folding dynamics of individual titin molecules in real time. We have also shown that fluorescence quenching can signal folding and unfolding of a small protein with only one immunoglobulin domain.

Bibliography

Zhuang, X., Ha, T., Kim, H. D., Centner, T., Labeit, S., & Chu, S. (2000). Fluorescence quenching: A tool for single-molecule protein-folding study. Proceedings of the National Academy of Sciences, 97(26), 14241â14244.

Authors 6

Xiaowei Zhuang (first)
Taekjip Ha (additional)
Harold D. Kim (additional)
Thomas Centner (additional)
Siegfried Labeit (additional)
Steven Chu (additional)

References 30 Referenced 175

10.1038/369183a0
10.1146/annurev.physchem.48.1.545
10.1038/nsb0197-10
10.1126/science.288.5473.2048
10.1073/pnas.090104997
10.1126/science.276.5315.1109
10.1038/30270
10.1126/science.288.5463.143
10.1126/science.276.5315.1112
10.1038/387308a0
10.1006/abio.1997.2259
J N Weinstein, E Ralston, L D Leserman, R D Klausner, P Dragsten, P Henkart, R Blumenthal Liposomen Technology, ed G Gregoriadis (CRC, Boca Raton, FL), pp. 183–204 (1984). / Liposomen Technology by Weinstein J N (1984)
10.1021/bi00012a028
10.1126/science.270.5234.293
10.1083/jcb.106.5.1563
10.1002/j.1460-2075.1992.tb05222.x
10.1126/science.4071053
10.1016/S0006-3495(95)80278-X
10.1016/S0006-3495(95)80131-1
10.1161/01.RES.84.11.1339
10.1083/jcb.146.3.631
10.1006/jmbi.1996.0050
10.1103/PhysRevLett.49.394
10.1103/PhysRevB.34.7578
10.1103/PhysRevB.26.4836
10.1002/pssb.2220860202
Agranovich V. M. & Galanin M. D. Electronic Excitation Energy Transfer in Condensed Matter (North–Holland Amsterdam).
10.1016/0003-2697(88)90412-5
B Stevens Adv Photochem 8, 161–226 (1971). (10.1002/9780470133385.ch4) / Adv Photochem by Stevens B (1971)
10.1088/0034-4885/38/8/001

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:36 a.m.)
Deposited	1 year, 7 months ago (Jan. 3, 2024, 11:54 p.m.)
Indexed	8 hours, 20 minutes ago (Aug. 27, 2025, 12:12 p.m.)
Issued	24 years, 8 months ago (Dec. 19, 2000)
Published	24 years, 8 months ago (Dec. 19, 2000)
Published Online	24 years, 8 months ago (Dec. 19, 2000)
Published Print	24 years, 8 months ago (Dec. 19, 2000)

Funders 0

None

BibTeX

@article{Zhuang_2000, title={Fluorescence quenching: A tool for single-molecule protein-folding study}, volume={97}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.97.26.14241}, DOI={10.1073/pnas.97.26.14241}, number={26}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Zhuang, Xiaowei and Ha, Taekjip and Kim, Harold D. and Centner, Thomas and Labeit, Siegfried and Chu, Steven}, year={2000}, month=dec, pages={14241–14244} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        8,
        27
      ]
    ],
    "date-time": "2025-08-27T16:12:31Z",
    "timestamp": 1756311151079
  },
  "reference-count": 30,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "26",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2000,
        12,
        19
      ]
    ]
  },
  "abstract": "<jats:p>By using titin as a model system, we have demonstrated that fluorescence quenching can be used to study protein folding at the single molecule level. The unfolded titin molecules with multiple dye molecules attached are able to fold to the native state. In the native folded state, the fluorescence from dye molecules is quenched due to the close proximity between the dye molecules. Unfolding of the titin leads to a dramatic increase in the fluorescence intensity. Such a change makes the folded and unfolded states of a single titin molecule clearly distinguishable and allows us to measure the folding dynamics of individual titin molecules in real time. We have also shown that fluorescence quenching can signal folding and unfolding of a small protein with only one immunoglobulin domain.</jats:p>",
  "DOI": "10.1073/pnas.97.26.14241",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        26
      ]
    ],
    "date-time": "2002-07-26T14:36:44Z",
    "timestamp": 1027694204000
  },
  "page": "14241-14244",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 175,
  "title": "Fluorescence quenching: A tool for single-molecule protein-folding study",
  "prefix": "10.1073",
  "volume": "97",
  "author": [
    {
      "given": "Xiaowei",
      "family": "Zhuang",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Department of Physics, Stanford University, Stanford, CA 94305; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, P. O. Box 102209, 69012 Heidelberg, Germany; and Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA 99164"
        }
      ]
    },
    {
      "given": "Taekjip",
      "family": "Ha",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Physics, Stanford University, Stanford, CA 94305; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, P. O. Box 102209, 69012 Heidelberg, Germany; and Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA 99164"
        }
      ]
    },
    {
      "given": "Harold D.",
      "family": "Kim",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Physics, Stanford University, Stanford, CA 94305; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, P. O. Box 102209, 69012 Heidelberg, Germany; and Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA 99164"
        }
      ]
    },
    {
      "given": "Thomas",
      "family": "Centner",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Physics, Stanford University, Stanford, CA 94305; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, P. O. Box 102209, 69012 Heidelberg, Germany; and Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA 99164"
        }
      ]
    },
    {
      "given": "Siegfried",
      "family": "Labeit",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Physics, Stanford University, Stanford, CA 94305; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, P. O. Box 102209, 69012 Heidelberg, Germany; and Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA 99164"
        }
      ]
    },
    {
      "given": "Steven",
      "family": "Chu",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Physics, Stanford University, Stanford, CA 94305; European Molecular Biology Laboratory Heidelberg, Meyerhofstrasse 1, P. O. Box 102209, 69012 Heidelberg, Germany; and Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA 99164"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        2000,
        12,
        19
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_2_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/369183a0"
    },
    {
      "key": "e_1_3_2_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.physchem.48.1.545"
    },
    {
      "key": "e_1_3_2_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nsb0197-10"
    },
    {
      "key": "e_1_3_2_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.288.5473.2048"
    },
    {
      "key": "e_1_3_2_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.090104997"
    },
    {
      "key": "e_1_3_2_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.276.5315.1109"
    },
    {
      "key": "e_1_3_2_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/30270"
    },
    {
      "key": "e_1_3_2_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.288.5463.143"
    },
    {
      "key": "e_1_3_2_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.276.5315.1112"
    },
    {
      "key": "e_1_3_2_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/387308a0"
    },
    {
      "key": "e_1_3_2_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/abio.1997.2259"
    },
    {
      "key": "e_1_3_2_12_2",
      "first-page": "183",
      "volume-title": "Liposomen Technology",
      "author": "Weinstein J N",
      "year": "1984",
      "unstructured": "J N Weinstein, E Ralston, L D Leserman, R D Klausner, P Dragsten, P Henkart, R Blumenthal Liposomen Technology, ed G Gregoriadis (CRC, Boca Raton, FL), pp. 183\u2013204 (1984)."
    },
    {
      "key": "e_1_3_2_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00012a028"
    },
    {
      "key": "e_1_3_2_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.270.5234.293"
    },
    {
      "key": "e_1_3_2_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.106.5.1563"
    },
    {
      "key": "e_1_3_2_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/j.1460-2075.1992.tb05222.x"
    },
    {
      "key": "e_1_3_2_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.4071053"
    },
    {
      "key": "e_1_3_2_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0006-3495(95)80278-X"
    },
    {
      "key": "e_1_3_2_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0006-3495(95)80131-1"
    },
    {
      "key": "e_1_3_2_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1161/01.RES.84.11.1339"
    },
    {
      "key": "e_1_3_2_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.146.3.631"
    },
    {
      "key": "e_1_3_2_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.1996.0050"
    },
    {
      "key": "e_1_3_2_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1103/PhysRevLett.49.394"
    },
    {
      "key": "e_1_3_2_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1103/PhysRevB.34.7578"
    },
    {
      "key": "e_1_3_2_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1103/PhysRevB.26.4836"
    },
    {
      "key": "e_1_3_2_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/pssb.2220860202"
    },
    {
      "key": "e_1_3_2_27_2",
      "unstructured": "Agranovich V. M. & Galanin M. D. Electronic Excitation Energy Transfer in Condensed Matter (North\u2013Holland Amsterdam)."
    },
    {
      "key": "e_1_3_2_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0003-2697(88)90412-5"
    },
    {
      "key": "e_1_3_2_29_2",
      "doi-asserted-by": "crossref",
      "first-page": "161",
      "DOI": "10.1002/9780470133385.ch4",
      "volume": "8",
      "author": "Stevens B",
      "year": "1971",
      "unstructured": "B Stevens Adv Photochem 8, 161\u2013226 (1971).",
      "journal-title": "Adv Photochem"
    },
    {
      "key": "e_1_3_2_30_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1088/0034-4885/38/8/001"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.97.26.14241",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2024,
        1,
        4
      ]
    ],
    "date-time": "2024-01-04T04:54:08Z",
    "timestamp": 1704344048000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.97.26.14241"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2000,
        12,
        19
      ]
    ]
  },
  "references-count": 30,
  "journal-issue": {
    "issue": "26",
    "published-print": {
      "date-parts": [
        [
          2000,
          12,
          19
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.97.26.14241"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.97.26.14241",
  "relation": {},
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        2000,
        12,
        19
      ]
    ]
  },
  "assertion": [
    {
      "value": "2000-10-30",
      "order": 1,
      "name": "accepted",
      "label": "Accepted",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    },
    {
      "value": "2000-12-19",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}