title: Single-molecule force spectroscopy approach to enzyme catalysis.
abstract: Enzyme catalysis has been traditionally studied using a diverse set of techniques such as bulk biochemistry, x-ray crystallography, and NMR. Recently, single-molecule force spectroscopy by atomic force microscopy has been used as a new tool to study the catalytic properties of an enzyme. In this approach, a mechanical force ranging up to hundreds of piconewtons is applied to the substrate of an enzymatic reaction, altering the conformational energy of the substrate-enzyme interactions during catalysis. From these measurements, the force dependence of an enzymatic reaction can be determined. The force dependence provides valuable new information about the dynamics of enzyme catalysis with sub-angstrom resolution, a feat unmatched by any other current technique. To date, single-molecule force spectroscopy has been applied to gain insight into the reduction of disulfide bonds by different enzymes of the thioredoxin family. This minireview aims to present a perspective on this new approach to study enzyme catalysis and to summarize the results that have already been obtained from it. Finally, the specific requirements that must be fulfilled to apply this new methodology to any other enzyme will be discussed.
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027, USA.
authors.forename: Jorge
authors.initials: J
authors.lastname: Alegre-Cebollada
authors.forename: Raul
authors.initials: R
authors.lastname: Perez-Jimenez
authors.forename: Pallav
authors.initials: P
authors.lastname: Kosuri
authors.forename: Julio M
authors.initials: JM
authors.lastname: Fernandez
mesh_terms: D001703:Biophysics / Q000379:methods; D002384:Catalysis; D004220:Disulfides / Q000737:chemistry; D004798:Enzymes / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D018625:Microscopy, Atomic Force / Q000379:methods*; D008968:Molecular Conformation; D010100:Oxygen / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D013053:Spectrophotometry / Q000379:methods; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013879:Thioredoxins / Q000737:chemistry
chemical_list: D004220:Disulfides; D004798:Enzymes; D011506:Proteins; D013438:Sulfhydryl Compounds; D013879:Thioredoxins; D010100:Oxygen
references.citation: Nature. 1978 Sep 21;275(5677):247-8
references.pmid: 692702
references.citation: Science. 2003 Dec 19;302(5653):2130-4
references.pmid: 14657506
references.citation: Chem Rev. 2004 Aug;104(8):3623-40
references.pmid: 15303831
references.citation: J Biol Chem. 1972 Apr 10;247(7):1992-8
references.pmid: 4552684
references.citation: Proc Natl Acad Sci U S A. 2001 Jan 16;98(2):468-72
references.pmid: 11149943
references.citation: Annu Rev Biochem. 2003;72:517-71
references.pmid: 12704087
references.citation: Biochemistry. 2008 Mar 18;47(11):3317-21
references.pmid: 18298083
references.citation: J Biol Chem. 1979 Oct 10;254(19):9627-32
references.pmid: 385588
references.citation: J Biol Chem. 1979 Sep 25;254(18):9113-9
references.pmid: 39074
references.citation: Biophys J. 2007 Jan 1;92(1):225-33
references.pmid: 17028145
references.citation: Annu Rev Biochem. 2006;75:519-41
references.pmid: 16756501
references.citation: J Am Chem Soc. 2008 May 21;130(20):6479-87
references.pmid: 18433129
references.citation: Langmuir. 2008 Feb 19;24(4):1356-64
references.pmid: 17999545
references.citation: Structure. 2005 Jun;13(6):893-904
references.pmid: 15939021
references.citation: Biochemistry. 2005 Dec 27;44(51):16835-43
references.pmid: 16363797
references.citation: Science. 2006 Sep 15;313(5793):1638-42
references.pmid: 16973882
references.citation: J Biol Chem. 1991 May 25;266(15):9494-500
references.pmid: 2033048
references.citation: Science. 1999 Mar 12;283(5408):1727-30
references.pmid: 10073936
references.citation: Chem Rev. 2006 May;106(5):1737-56
references.pmid: 16683752
references.citation: Proc Natl Acad Sci U S A. 2004 May 11;101(19):7299-304
references.pmid: 15123816
references.citation: Curr Opin Chem Biol. 2007 Apr;11(2):174-81
references.pmid: 17353141
references.citation: Antioxid Redox Signal. 2007 Jan;9(1):25-47
references.pmid: 17115886
references.citation: Annu Rev Biochem. 1985;54:237-71
references.pmid: 3896121
references.citation: Nature. 2007 Nov 1;450(7166):124-7
references.pmid: 17972886
references.citation: J Biol Chem. 2008 Oct 3;283(40):27121-9
references.pmid: 18687682
references.citation: Chemistry. 2004 Jan 5;10(1):257-66
references.pmid: 14695571
references.citation: Nature. 2007 Dec 6;450(7171):913-6
references.pmid: 18026087
references.citation: Nature. 2005 Nov 3;438(7064):117-21
references.pmid: 16267559
references.citation: Annu Rev Biochem. 1983;52:263-300
references.pmid: 6351724
references.citation: Proc Natl Acad Sci U S A. 2006 May 9;103(19):7222-7
references.pmid: 16645035
references.citation: Science. 1997 Jul 11;277(5323):202-6
references.pmid: 9211842
references.citation: Science. 1978 May 12;200(4342):618-27
references.pmid: 347575
references.citation: Curr Opin Struct Biol. 2005 Dec;15(6):604-13
references.pmid: 16274984
references.citation: Science. 2003 Aug 29;301(5637):1196-202
references.pmid: 12947189
references.citation: Chem Rev. 2005 Aug;105(8):2921-48
references.pmid: 16092823
references.citation: Nature. 2007 Nov 29;450(7170):750-4
references.pmid: 18004302
references.citation: Science. 1997 May 16;276(5315):1109-12
references.pmid: 9148804
references.citation: Biophys J. 2007 Oct 1;93(7):2436-46
references.pmid: 17545242
references.citation: Curr Opin Chem Biol. 2007 Apr;11(2):182-7
references.pmid: 17307018
references.citation: Nat Chem Biol. 2006 Feb;2(2):87-94
references.pmid: 16415859
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 24;103(43):15753-8
references.pmid: 17032759
references.citation: Methods Enzymol. 2005;394:507-24
references.pmid: 15808235
references.citation: Chemphyschem. 2008 Dec 22;9(18):2836-47
references.pmid: 19058277
references.citation: Nat Struct Mol Biol. 2009 Aug;16(8):890-6
references.pmid: 19597482
references.citation: J Am Chem Soc. 2008 Jan 16;130(2):436-7
references.pmid: 18088123
references.citation: Annu Rev Biophys. 2008;37:153-73
references.pmid: 18573077