DOI: 10.1073/pnas.0911085107. Structure of intact <i>Thermus thermophilus</i> V-ATPase by cryo-EM reveals organization of the membrane-bound V <sub>O</sub> motor

Structure of intact Thermus thermophilus V-ATPase by cryo-EM reveals organization of the membrane-bound V O motor

10.1073/pnas.0911085107

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

The eubacterium Thermus thermophilus uses a macromolecular assembly closely related to eukaryotic V-ATPase to produce its supply of ATP. This simplified V-ATPase offers several advantages over eukaryotic V-ATPases for structural analysis and investigation of the mechanism of the enzyme. Here we report the structure of the complex at ∼16 Å resolution as determined by single particle electron cryomicroscopy (cryo-EM). The resolution of the map and our use of cryo-EM, rather than negative stain EM, reveals detailed information about the internal organization of the assembly. We could separate the map into segments corresponding to subunits A and B, the threefold pseudosymmetric C-subunit, a central rotor consisting of subunits D and F, the L-ring, the stator subcomplex consisting of subunits I, E, and G, and a micelle of bound detergent. The architecture of the V O region shows a remarkably small area of contact between the I-subunit and the ring of L-subunits and is consistent with a two half-channel model for proton translocation. The arrangement of structural elements in V O gives insight into the mechanism of torque generation from proton translocation.

Bibliography

Lau, W. C. Y., & Rubinstein, J. L. (2010). Structure of intact Thermus thermophilus V-ATPase by cryo-EM reveals organization of the membrane-bound V O motor. Proceedings of the National Academy of Sciences, 107(4), 1367â1372.

Authors 2

Wilson C. Y. Lau (first)
John L. Rubinstein (additional)

References 57 Referenced 64

10.1007/s10863-007-9112-z
10.1038/nrm2272
10.1038/370621a0
10.1038/386299a0
10.1038/381623a0
10.1073/pnas.0436796100
10.1007/s10863-005-9480-1
10.1016/S0021-9258(17)46944-4
10.1074/jbc.270.10.5649
10.1074/jbc.M408278200
10.1016/j.jmb.2008.08.014
10.1093/emboj/cdg608
10.1074/jbc.M704941200
10.1073/pnas.0706914105
10.1074/jbc.M305853200
10.1016/j.str.2004.07.017
10.1016/j.str.2008.09.010
10.1074/jbc.M808498200
10.1074/jbc.M407821200
10.1074/jbc.M805345200
10.1016/j.jmb.2009.01.014
10.1038/emboj.2009.310
10.1038/embor.2009.202
10.1073/pnas.0305165101
10.1016/j.jmb.2006.11.088
10.1126/science.1110064
10.1038/sj.embor.7400294
10.1073/pnas.131192798
10.1146/annurev.biochem.66.1.717
10.1002/(SICI)1521-3773(19980918)37:17<2308::AID-ANIE2308>3.0.CO;2-W
10.1126/science.1112617
10.1016/S0968-0004(97)01129-8
10.1074/jbc.275.18.13955
10.1016/S0021-9258(18)94065-2
10.1073/pnas.221291798
10.1126/science.1111199
10.1016/j.jsb.2008.01.001
10.1016/j.jsb.2006.05.004
PB Rosenthal, RA Crowther, R Henderson, An objective criterion for resolution assessment in single-particle electron microscopy (appendix). J Mol Biol 333, 743–745 (2003). / J Mol Biol / An objective criterion for resolution assessment in single-particle electron microscopy (appendix) by Rosenthal PB (2003)
10.1016/j.jsb.2006.08.003
10.1016/j.jmb.2003.07.013
10.1016/0014-5793(88)80513-1
10.1038/314076a0
10.1016/j.ymeth.2006.07.019
10.1073/pnas.0607640103
10.1016/j.molcel.2007.10.034
10.1128/MMBR.70.1.177-191.2006
10.1126/science.286.5445.1700
10.1016/S0014-5793(01)02837-X
10.1074/jbc.273.32.20504
10.1074/jbc.M801276200
10.1006/jsbi.1999.4174
10.1146/annurev.biophys.29.1.291
10.1110/ps.062095806
10.1006/jmbi.2002.5438
10.1016/j.jsb.2006.06.010
10.1038/46224

Dates

Type	When
Created	15 years, 7 months ago (Jan. 6, 2010, 10:37 p.m.)
Deposited	3 years, 4 months ago (April 12, 2022, 6:50 p.m.)
Indexed	1 month, 2 weeks ago (July 2, 2025, 2:33 p.m.)
Issued	15 years, 7 months ago (Jan. 6, 2010)
Published	15 years, 7 months ago (Jan. 6, 2010)
Published Online	15 years, 7 months ago (Jan. 6, 2010)
Published Print	15 years, 6 months ago (Jan. 26, 2010)

Funders 0

None

BibTeX

@article{Lau_2010, title={Structure of intact Thermus thermophilus V-ATPase by cryo-EM reveals organization of the membrane-bound V O motor}, volume={107}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0911085107}, DOI={10.1073/pnas.0911085107}, number={4}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Lau, Wilson C. Y. and Rubinstein, John L.}, year={2010}, month=jan, pages={1367–1372} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        2
      ]
    ],
    "date-time": "2025-07-02T18:33:55Z",
    "timestamp": 1751481235551
  },
  "reference-count": 57,
  "publisher": "Proceedings of the National Academy of Sciences",
  "issue": "4",
  "content-domain": {
    "domain": [
      "www.pnas.org"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2010,
        1,
        26
      ]
    ]
  },
  "abstract": "<jats:p>\n            The eubacterium\n            <jats:italic>Thermus thermophilus</jats:italic>\n            uses a macromolecular assembly closely related to eukaryotic V-ATPase to produce its supply of ATP. This simplified V-ATPase offers several advantages over eukaryotic V-ATPases for structural analysis and investigation of the mechanism of the enzyme. Here we report the structure of the complex at \u223c16\u00a0\n            <jats:italic>\u212b</jats:italic>\n            resolution as determined by single particle electron cryomicroscopy (cryo-EM). The resolution of the map and our use of cryo-EM, rather than negative stain EM, reveals detailed information about the internal organization of the assembly. We could separate the map into segments corresponding to subunits A and B, the threefold pseudosymmetric C-subunit, a central rotor consisting of subunits D and F, the L-ring, the stator subcomplex consisting of subunits I, E, and G, and a micelle of bound detergent. The architecture of the V\n            <jats:sub>O</jats:sub>\n            region shows a remarkably small area of contact between the I-subunit and the ring of L-subunits and is consistent with a two half-channel model for proton translocation. The arrangement of structural elements in V\n            <jats:sub>O</jats:sub>\n            gives insight into the mechanism of torque generation from proton translocation.\n          </jats:p>",
  "DOI": "10.1073/pnas.0911085107",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2010,
        1,
        7
      ]
    ],
    "date-time": "2010-01-07T03:37:14Z",
    "timestamp": 1262835434000
  },
  "page": "1367-1372",
  "update-policy": "http://dx.doi.org/10.1073/pnas.cm10313",
  "source": "Crossref",
  "is-referenced-by-count": 64,
  "title": "Structure of intact\n            <i>Thermus thermophilus</i>\n            V-ATPase by cryo-EM reveals organization of the membrane-bound V\n            <sub>O</sub>\n            motor",
  "prefix": "10.1073",
  "volume": "107",
  "author": [
    {
      "given": "Wilson C. Y.",
      "family": "Lau",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Molecular Structure and Function Program, The Hospital for Sick Children Research Institute and Department of Biochemistry, University of Toronto, Toronto, Ontario M5G 1X8, Canada"
        }
      ]
    },
    {
      "given": "John L.",
      "family": "Rubinstein",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Molecular Structure and Function Program, The Hospital for Sick Children Research Institute and Department of Biochemistry, University of Toronto, Toronto, Ontario M5G 1X8, Canada"
        }
      ]
    }
  ],
  "member": "341",
  "published-online": {
    "date-parts": [
      [
        2010,
        1,
        6
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_3_3_1_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s10863-007-9112-z"
    },
    {
      "key": "e_1_3_3_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/nrm2272"
    },
    {
      "key": "e_1_3_3_3_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/370621a0"
    },
    {
      "key": "e_1_3_3_4_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/386299a0"
    },
    {
      "key": "e_1_3_3_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/381623a0"
    },
    {
      "key": "e_1_3_3_6_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0436796100"
    },
    {
      "key": "e_1_3_3_7_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s10863-005-9480-1"
    },
    {
      "key": "e_1_3_3_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(17)46944-4"
    },
    {
      "key": "e_1_3_3_9_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.270.10.5649"
    },
    {
      "key": "e_1_3_3_10_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M408278200"
    },
    {
      "key": "e_1_3_3_11_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jmb.2008.08.014"
    },
    {
      "key": "e_1_3_3_12_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/cdg608"
    },
    {
      "key": "e_1_3_3_13_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M704941200"
    },
    {
      "key": "e_1_3_3_14_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0706914105"
    },
    {
      "key": "e_1_3_3_15_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M305853200"
    },
    {
      "key": "e_1_3_3_16_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.str.2004.07.017"
    },
    {
      "key": "e_1_3_3_17_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.str.2008.09.010"
    },
    {
      "key": "e_1_3_3_18_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M808498200"
    },
    {
      "key": "e_1_3_3_19_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M407821200"
    },
    {
      "key": "e_1_3_3_20_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M805345200"
    },
    {
      "key": "e_1_3_3_21_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jmb.2009.01.014"
    },
    {
      "key": "e_1_3_3_22_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/emboj.2009.310"
    },
    {
      "key": "e_1_3_3_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/embor.2009.202"
    },
    {
      "key": "e_1_3_3_24_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0305165101"
    },
    {
      "key": "e_1_3_3_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jmb.2006.11.088"
    },
    {
      "key": "e_1_3_3_26_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1110064"
    },
    {
      "key": "e_1_3_3_27_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/sj.embor.7400294"
    },
    {
      "key": "e_1_3_3_28_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.131192798"
    },
    {
      "key": "e_1_3_3_29_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.biochem.66.1.717"
    },
    {
      "key": "e_1_3_3_30_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/(SICI)1521-3773(19980918)37:17<2308::AID-ANIE2308>3.0.CO;2-W"
    },
    {
      "key": "e_1_3_3_31_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1112617"
    },
    {
      "key": "e_1_3_3_32_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0968-0004(97)01129-8"
    },
    {
      "key": "e_1_3_3_33_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.275.18.13955"
    },
    {
      "key": "e_1_3_3_34_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0021-9258(18)94065-2"
    },
    {
      "key": "e_1_3_3_35_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.221291798"
    },
    {
      "key": "e_1_3_3_36_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1111199"
    },
    {
      "key": "e_1_3_3_37_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jsb.2008.01.001"
    },
    {
      "key": "e_1_3_3_38_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jsb.2006.05.004"
    },
    {
      "key": "e_1_3_3_39_2",
      "first-page": "743",
      "article-title": "An objective criterion for resolution assessment in single-particle electron microscopy (appendix)",
      "volume": "333",
      "author": "Rosenthal PB",
      "year": "2003",
      "unstructured": "PB Rosenthal, RA Crowther, R Henderson, An objective criterion for resolution assessment in single-particle electron microscopy (appendix). J Mol Biol 333, 743\u2013745 (2003).",
      "journal-title": "J Mol Biol"
    },
    {
      "key": "e_1_3_3_40_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jsb.2006.08.003"
    },
    {
      "key": "e_1_3_3_41_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jmb.2003.07.013"
    },
    {
      "key": "e_1_3_3_42_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0014-5793(88)80513-1"
    },
    {
      "key": "e_1_3_3_43_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/314076a0"
    },
    {
      "key": "e_1_3_3_44_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.ymeth.2006.07.019"
    },
    {
      "key": "e_1_3_3_45_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.0607640103"
    },
    {
      "key": "e_1_3_3_46_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.molcel.2007.10.034"
    },
    {
      "key": "e_1_3_3_47_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1128/MMBR.70.1.177-191.2006"
    },
    {
      "key": "e_1_3_3_48_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.286.5445.1700"
    },
    {
      "key": "e_1_3_3_49_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0014-5793(01)02837-X"
    },
    {
      "key": "e_1_3_3_50_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.273.32.20504"
    },
    {
      "key": "e_1_3_3_51_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1074/jbc.M801276200"
    },
    {
      "key": "e_1_3_3_52_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jsbi.1999.4174"
    },
    {
      "key": "e_1_3_3_53_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1146/annurev.biophys.29.1.291"
    },
    {
      "key": "e_1_3_3_54_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1110/ps.062095806"
    },
    {
      "key": "e_1_3_3_55_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmbi.2002.5438"
    },
    {
      "key": "e_1_3_3_56_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/j.jsb.2006.06.010"
    },
    {
      "key": "e_1_3_3_57_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/46224"
    }
  ],
  "container-title": "Proceedings of the National Academy of Sciences",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://pnas.org/doi/pdf/10.1073/pnas.0911085107",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2022,
        4,
        12
      ]
    ],
    "date-time": "2022-04-12T22:50:10Z",
    "timestamp": 1649803810000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://pnas.org/doi/full/10.1073/pnas.0911085107"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2010,
        1,
        6
      ]
    ]
  },
  "references-count": 57,
  "journal-issue": {
    "issue": "4",
    "published-print": {
      "date-parts": [
        [
          2010,
          1,
          26
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1073/pnas.0911085107"
  ],
  "URL": "http://dx.doi.org/10.1073/pnas.0911085107",
  "relation": {
    "has-review": [
      {
        "id-type": "doi",
        "id": "10.3410/f.2178956.1788054",
        "asserted-by": "object"
      }
    ]
  },
  "ISSN": [
    "0027-8424",
    "1091-6490"
  ],
  "subject": [],
  "container-title-short": "Proc. Natl. Acad. Sci. U.S.A.",
  "published": {
    "date-parts": [
      [
        2010,
        1,
        6
      ]
    ]
  },
  "assertion": [
    {
      "value": "2010-01-06",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}