Robert H. Fillingame

Position title: Professor Emeritus


Phone: (608) 262-1439

6204A Biochemical Sciences Building
440 Henry Mall, Madison, WI 53706

Robert Fillingame

• BS 1968, Washington State University
• PhD 1973, University of Washington, Seattle
• Postdoctoral, 1973-75, Harvard Medical School (E.P. Kennedy)

Honors & Awards

• Medical Alumni Named Professorship, 1988
• NIH Merit Award, 1994
• WARF Mid-Career Faculty Researcher Award, 1998
• Dean’s Teaching Award (School of Medicine), 2008
Chancellor’s Teaching Award , 2012

Research Interests

My research focuses on the molecular mechanism of ATP synthesis during oxidative phosphorylation. ATP synthesis is driven by a trans membrane proton gradient generated by the electron transport system. The enzyme catalyzing ATP synthesis is biologically unique in using the electrochemical energy of the proton gradient to drive the rotation of a molecular motor which in turn drives ATP synthesis. The ATP synthase has been termed the “world’s smallest rotary motor”. The process works as follows. Proton transport through the Fo membrane sector of the enzyme is coupled with the rotation of a molecular rotor composed of multiple c subunits. The c oligomeric rotor in turn drives ATP synthesis via the rotation of two attached subunits (gamma and epsilon) which cycle between circularly arranged multiple catalytic sites in the extramembranous F1 sector of the enzyme. In the complete process of ATP synthesis the electrochemical energy of the proton gradient is converted into mechanical energy and, via structural/mechanical changes in the catalytic sites, ultimately into the chemical energy of ATP. Our long term goal is to elucidate the structure and molecular mechanism of this fascinating machine.

Publications of Note

Perform a customized PubMed literature search for Dr. Fillingame.

• Steed PR, Kraft KA, Fillingame RH. Interacting cytoplasmic loops of subunits a
and c of Escherichia coli F1F0 ATP synthase gate H+ transport to the cytoplasm.
Proc Natl Acad Sci U S A. 2014 Nov 25;111(47):16730-5. doi:
10.1073/pnas.1414660111. Epub 2014 Nov 10. PubMed PMID: 25385585; PubMed Central
PMCID: PMC4250104.

• Fillingame RH, Steed PR. Half channels mediating H(+) transport and the
mechanism of gating in the Fo sector of Escherichia coli F1Fo ATP synthase.
Biochim Biophys Acta. 2014 Jul;1837(7):1063-8. doi: 10.1016/j.bbabio.2014.03.005.
Epub 2014 Mar 17. Review. PubMed PMID: 24650630.

• Steed PR, Fillingame RH. Residues in the polar loop of subunit c in
Escherichia coli ATP synthase function in gating proton transport to the
cytoplasm. J Biol Chem. 2014 Jan 24;289(4):2127-38. doi: 10.1074/jbc.M113.527879.
Epub 2013 Dec 2. PubMed PMID: 24297166; PubMed Central PMCID: PMC3900959.

• Moore KJ, Fillingame RH. Obstruction of transmembrane helical movements in
subunit a blocks proton pumping by F1Fo ATP synthase. J Biol Chem. 2013 Aug
30;288(35):25535-41. doi: 10.1074/jbc.M113.496794. Epub 2013 Jul 17. PubMed PMID:
23864659; PubMed Central PMCID: PMC3757214.

• DeLeon-Rangel J, Ishmukhametov RR, Jiang W, Fillingame RH, Vik SB.
Interactions between subunits a and b in the rotary ATP synthase as determined by
cross-linking. FEBS Lett. 2013 Apr 2;587(7):892-7. doi:
10.1016/j.febslet.2013.02.012. Epub 2013 Feb 14. PubMed PMID: 23416299; PubMed
Central PMCID: PMC3608841.

• Uhlemann EM, Pierson HE, Fillingame RH, Dmitriev OY. Cell-free synthesis of
membrane subunits of ATP synthase in phospholipid bicelles: NMR shows subunit a
fold similar to the protein in the cell membrane. Protein Sci. 2012
Feb;21(2):279-88. doi: 10.1002/pro.2014. Epub 2012 Jan 4. PubMed PMID: 22162071;
PubMed Central PMCID: PMC3324772.

• Dong H, Fillingame RH. Chemical reactivities of cysteine substitutions in
subunit a of ATP synthase define residues gating H+ transport from each side of
the membrane. J Biol Chem. 2010 Dec 17;285(51):39811-8. doi:
10.1074/jbc.M110.175844. Epub 2010 Oct 13. PubMed PMID: 20943664; PubMed Central
PMCID: PMC3000962.

• Steed PR, Fillingame RH. Aqueous accessibility to the transmembrane regions of
subunit c of the Escherichia coli F1F0 ATP synthase. J Biol Chem. 2009 Aug
28;284(35):23243-50. doi: 10.1074/jbc.M109.002501. Epub 2009 Jun 19. PubMed PMID:
19542218; PubMed Central PMCID: PMC2749098.

• Moore KJ, Fillingame RH. Structural interactions between transmembrane helices
4 and 5 of subunit a and the subunit c ring of Escherichia coli ATP synthase. J
Biol Chem
. 2008 Nov 14;283(46):31726-35. doi: 10.1074/jbc.M803848200. Epub 2008
Sep 11. PubMed PMID: 18786930; PubMed Central PMCID: PMC2581582.

10: Moore KJ, Angevine CM, Vincent OD, Schwem BE, Fillingame RH. The cytoplasmic
loops of subunit a of Escherichia coli ATP synthase may participate in the proton
translocating mechanism. J Biol Chem. 2008 May 9;283(19):13044-52. doi:
10.1074/jbc.M800900200. Epub 2008 Mar 12. PubMed PMID: 18337242; PubMed Central
PMCID: PMC2442323.

• Steed PR, Fillingame RH. Subunit a facilitates aqueous access to a
membrane-embedded region of subunit c in Escherichia coli F1F0 ATP synthase. J
Biol Chem
. 2008 May 2;283(18):12365-72. doi: 10.1074/jbc.M800901200. Epub 2008
Mar 10. PubMed PMID: 18332132; PubMed Central PMCID: PMC2431001.

• Dmitriev OY, Freedman KH, Hermolin J, Fillingame RH. Interaction of
transmembrane helices in ATP synthase subunit a in solution as revealed by spin
label difference NMR. Biochim Biophys Acta. 2008 Feb;1777(2):227-37. doi:
10.1016/j.bbabio.2007.11.011. Epub 2007 Dec 15. PubMed PMID: 18178144; PubMed
Central PMCID: PMC2276983.

• Vincent OD, Schwem BE, Steed PR, Jiang W, Fillingame RH. Fluidity of
structure and swiveling of helices in the subunit c ring of Escherichia coli ATP
synthase as revealed by cysteine-cysteine cross-linking. J Biol Chem. 2007 Nov
16;282(46):33788-94. Epub 2007 Sep 24. PubMed PMID: 17893141.

• Dmitriev OY, Fillingame RH. The rigid connecting loop stabilizes hairpin
folding of the two helices of the ATP synthase subunit c. Protein Sci. 2007
Oct;16(10):2118-22. Epub 2007 Aug 31. PubMed PMID: 17766379; PubMed Central
PMCID: PMC2204134.

• Angevine CM, Herold KA, Vincent OD, Fillingame RH. Aqueous access pathways in
ATP synthase subunit a. Reactivity of cysteine substituted into transmembrane
helices 1, 3, and 5. J Biol Chem. 2007 Mar 23;282(12):9001-7. Epub 2007 Jan 18.
PubMed PMID: 17234633.

• Schwem BE, Fillingame RH. Cross-linking between helices within subunit a of
Escherichia coli ATP synthase defines the transmembrane packing of a four-helix
bundle. J Biol Chem. 2006 Dec 8;281(49):37861-7. Epub 2006 Oct 11. PubMed PMID:

• Fillingame RH, Jiang W, Dmitriev OY. The oligomeric subunit C rotor in the fo
sector of ATP synthase: unresolved questions in our understanding of function. J
Bioenerg Biomembr
. 2000 Oct;32(5):433-9. PubMed PMID: 15254378.

• Dmitriev OY, Abildgaard F, Markley JL, Fillingame RH. Backbone 1H, 15N and
13C assignments for the subunit a of the E. coli ATP synthase. J Biomol NMR. 2004
Jul;29(3):439-40. PubMed PMID: 15213458.

• Aksimentiev A, Balabin IA, Fillingame RH, Schulten K. Insights into the
molecular mechanism of rotation in the Fo sector of ATP synthase. Biophys J. 2004
Mar;86(3):1332-44. PubMed PMID: 14990464; PubMed Central PMCID: PMC1303972.

• Dmitriev OY, Altendorf K, Fillingame RH. Subunit A of the E. coli ATP
synthase: reconstitution and high resolution NMR with protein purified in a mixed
polarity solvent. FEBS Lett. 2004 Jan 2;556(1-3):35-8. PubMed PMID: 14706821.