title: Dodine as a protein denaturant: the best of two worlds?
abstract: Traditional denaturants such as urea and guanidinium ion unfold proteins in a cooperative "all-or-none" fashion. However, their high working concentration in combination with their strong absorption in the far ultraviolet region make it impossible to measure high quality circular dichroism or infrared spectra, which are commonly used to detect changes in protein secondary structure. On the other hand, detergents such as dodecyl sulfate destabilize native protein conformation at low millimolar concentrations and are UV transparent, but they denature proteins more gradually than guanidinium or urea. In this work, we studied the denaturation properties of the fungicide dodecylguanidinium acetate (dodine), which combines both denaturants into one. We show that dodine unfolds some small proteins at millimolar concentrations, facilitates temperature denaturation, and is transparent enough at its working concentration, unlike guanidinium, to measure full range circular dichroism spectra. Our results also suggest that dodine allows fine-tuning of the protein's unfolded state, unlike traditional "all-or-none" denaturants.
authors.affiliation: Department of Physics, Center for the Physics of Living Cells, University of Illinois , Urbana, Illinois 61801, United States.
authors.forename: Hannah
authors.initials: H
authors.lastname: Gelman
authors.forename: Tatyana
authors.initials: T
authors.lastname: Perlova
authors.forename: Martin
authors.initials: M
authors.lastname: Gruebele
mesh_terms: D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry; D006146:Guanidines / Q000737:chemistry*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D014508:Urea / Q000737:chemistry; D054334:Viral Regulatory and Accessory Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D006146:Guanidines; D011506:Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins; C006921:dodine; D012967:Sodium Dodecyl Sulfate; D014508:Urea; C028913:dodecyl sulfate; D019791:Guanidine
references.citation: J Am Chem Soc. 2011 Dec 7;133(48):19338-41
references.pmid: 22066714
references.citation: Nature. 2003 May 8;423(6936):193-7
references.pmid: 12736690
references.citation: Biochim Biophys Acta. 2011 May;1814(5):562-91
references.pmid: 21397738
references.citation: J Mol Biol. 2007 Jul 13;370(3):574-84
references.pmid: 17532338
references.citation: Biochim Biophys Acta. 1983 Feb 28;743(1):31-6
references.pmid: 6600627
references.citation: J Biol Chem. 1974 Sep 10;249(17):5388-93
references.pmid: 4416801
references.citation: Annu Rev Biochem. 1991;60:795-825
references.pmid: 1883209
references.citation: PLoS One. 2010 Apr 21;5(4):e10262
references.pmid: 20421996
references.citation: J Mol Biol. 2002 Apr 5;317(4):613-27
references.pmid: 11955013
references.citation: J Biol Chem. 2001 Aug 31;276(35):32403-6
references.pmid: 11432878
references.citation: Biophys J. 2007 May 15;92(10):3674-85
references.pmid: 17351005
references.citation: Proc Natl Acad Sci U S A. 2013 May 14;110(20):8087-92
references.pmid: 23620522
references.citation: Proc Natl Acad Sci U S A. 2008 Jan 8;105(1):118-23
references.pmid: 18172203
references.citation: J Am Chem Soc. 2004 Nov 17;126(45):14686-7
references.pmid: 15535670
references.citation: Nature. 2002 Nov 7;420(6911):102-6
references.pmid: 12422224
references.citation: Biophys J. 2002 Oct;83(4):2219-30
references.pmid: 12324439
references.citation: Biochemistry. 1966 Apr;5(4):1242-54
references.pmid: 5958200
references.citation: J Mol Biol. 2011 Jan 7;405(1):43-8
references.pmid: 20974152
references.citation: J Phys Chem B. 2011 Mar 10;115(9):2090-6
references.pmid: 21319829
references.citation: J Chem Phys. 2009 Nov 21;131(19):195101
references.pmid: 19929078
references.citation: Science. 2011 Oct 28;334(6055):517-20
references.pmid: 22034434
references.citation: Proc Natl Acad Sci U S A. 2009 Sep 22;106(38):16239-44
references.pmid: 19805287
references.citation: Biochemistry. 2005 Aug 2;44(30):10406-15
references.pmid: 16042418
references.citation: Biophys J. 2009 Sep 16;97(6):1772-7
references.pmid: 19751683
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2369-74
references.pmid: 18268349
references.citation: J Mol Biol. 2010 Apr 2;397(3):789-98
references.pmid: 20138892
references.citation: Adv Protein Chem. 1968;23:121-282
references.pmid: 4882248
references.citation: Biochemistry. 1968 Nov;7(11):4135-8
references.pmid: 5749177
references.citation: Annu Rev Phys Chem. 2011;62:257-77
references.pmid: 21219136