title: Reverse micelles as a tool for probing solvent modulation of protein dynamics: Reverse micelle encapsulated hemoglobin.
abstract: Hydration waters impact protein dynamics. Dissecting the interplay between hydration waters and dynamics requires a protein that manifests a broad range of dynamics. Proteins in reverse micelles (RMs) have promise as tools to achieve this objective because the water content can be manipulated. Hemoglobin is an appropriate tool with which to probe hydration effects. We describe both a protocol for hemoglobin encapsulation in reverse micelles and a facile method using PEG and cosolvents to manipulate water content. Hydration properties are probed using the water-sensitive fluorescence from Hb bound pyranine and covalently attached Badan. Protein dynamics are probed through ligand recombination traces derived from photodissociated carbonmonoxy hemoglobin on a log scale that exposes the potential role of both α and β solvent fluctuations in modulating protein dynamics. The results open the possibility of probing hydration level phenomena in this system using a combination of NMR and optical probes.
authors.affiliation: Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, United States.
authors.forename: Camille J
authors.initials: CJ
authors.lastname: Roche
authors.forename: David
authors.initials: D
authors.lastname: Dantsker
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Heller
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Sabat
authors.forename: Joel M
authors.initials: JM
authors.lastname: Friedman
keywords: Geminate recombination; Hemoglobin; Hydration; Protein dynamics; Reverse micelles; Solvent slaving
references.citation: J Biol Chem. 1983 Feb 25;258(4):2296-302
references.pmid: 6822561
references.citation: Biophys Chem. 2002 Jul 10;98(1-2):35-48
references.pmid: 12128188
references.citation: J Biomol NMR. 1999 May;14(1):75-8
references.pmid: 10382308
references.citation: J Biol Chem. 2002 Jul 12;277(28):25783-90
references.pmid: 11976324
references.citation: J Am Chem Soc. 2007 Oct 24;129(42):12756-64
references.pmid: 17910446
references.citation: Phys Rev Lett. 2005 Feb 11;94(5):057405
references.pmid: 15783696
references.citation: Biopolymers. 2004 May-Jun 5;74(1-2):60-3
references.pmid: 15137095
references.citation: Biochem Biophys Res Commun. 2004 Oct 22;323(3):760-8
references.pmid: 15381065
references.citation: J Biol Chem. 2006 Nov 24;281(47):36021-8
references.pmid: 17005567
references.citation: J Biol Chem. 2005 Nov 18;280(46):38740-55
references.pmid: 16155005
references.citation: Eur Biophys J. 2010 Mar;39(4):647-56
references.pmid: 19760185
references.citation: J Am Chem Soc. 2012 May 23;134(20):8543-50
references.pmid: 22452540
references.citation: Biochemistry. 2004 Apr 27;43(16):4832-43
references.pmid: 15096052
references.citation: J Biol Chem. 1981 Jan 25;256(2):837-43
references.pmid: 7192710
references.citation: Biochim Biophys Acta. 2012 May;1824(5):769-84
references.pmid: 22446163
references.citation: J Phys Chem B. 2011 Oct 13;115(40):11658-70
references.pmid: 21899355
references.citation: Biochemistry. 1998 Jul 14;37(28):9940-51
references.pmid: 9665699
references.citation: Chem Rev. 2006 May;106(5):1672-99
references.pmid: 16683749
references.citation: J Phys Chem B. 2005 Nov 17;109(45):21273-84
references.pmid: 16853758
references.citation: J Am Chem Soc. 2006 Aug 23;128(33):10652-3
references.pmid: 16910639
references.citation: Biophys J. 2008 Jul;95(1):322-32
references.pmid: 18339762
references.citation: Eur Biophys J. 2008 Jun;37(5):591-602
references.pmid: 18270694
references.citation: J Am Chem Soc. 2011 Aug 17;133(32):12326-9
references.pmid: 21761828
references.citation: J Colloid Interface Sci. 2010 Jun 15;346(2):391-7
references.pmid: 20371070
references.citation: IUBMB Life. 2007 Aug-Sep;59(8-9):506-12
references.pmid: 17701545
references.citation: Biochemistry. 1994 Nov 8;33(44):13132-9
references.pmid: 7947719
references.citation: J Am Chem Soc. 2009 Aug 12;131(31):11010-8
references.pmid: 19603752
references.citation: Biophys J. 2008 May 15;94(10):3945-55
references.pmid: 18234831
references.citation: Biophys J. 2007 Mar 15;92(6):L43-5
references.pmid: 17218465
references.citation: Biochemistry. 2001 Apr 3;40(13):3780-95
references.pmid: 11300758
references.citation: J Am Chem Soc. 2008 Oct 22;130(42):13927-37
references.pmid: 18823116
references.citation: Chirality. 2000 May;12(4):216-20
references.pmid: 10790192
references.citation: Eur J Biochem. 1982 Jan;121(3):637-42
references.pmid: 6276178
references.citation: J Phys Chem B. 2005 Apr 21;109(15):7488-99
references.pmid: 16851860
references.citation: J Biol Chem. 2006 Dec 15;281(50):38757-68
references.pmid: 17057250
references.citation: Biotechnol Prog. 1996 Nov-Dec;12(6):793-800
references.pmid: 8983206
references.citation: J Chem Phys. 2005 May 1;122(17):174501
references.pmid: 15910039
references.citation: J Phys Chem B. 2008 Aug 21;112(33):10158-64
references.pmid: 18651765
references.citation: J Inorg Biochem. 2000 Nov;82(1-4):93-101
references.pmid: 11132645
references.citation: J Am Chem Soc. 2012 Jul 25;134(29):11904-7
references.pmid: 22765228
references.citation: J Phys Chem A. 2011 Jan 20;115(2):128-35
references.pmid: 21155585
references.citation: Biophys J. 1999 Oct;77(4):2237-50
references.pmid: 10512843
references.citation: Biochim Biophys Acta. 2005 Jun 1;1749(2):234-51
references.pmid: 15914102
references.citation: Biophys J. 2009 Oct 21;97(8):2258-66
references.pmid: 19843458
references.citation: J Biol Chem. 1969 Mar 25;244(6):1651-3
references.pmid: 5773062
references.citation: Protein Eng. 1998 Jul;11(7):583-8
references.pmid: 9740377
references.citation: J Phys Chem B. 2011 Apr 14;115(14):4099-109
references.pmid: 21425816
references.citation: J Phys Chem B. 2009 Dec 31;113(52):16632-42
references.pmid: 19961206
references.citation: J Am Chem Soc. 2007 Jul 4;129(26):8122-30
references.pmid: 17567012
references.citation: Biochemistry. 2006 Jul 25;45(29):8732-41
references.pmid: 16846216
references.citation: J Biomol NMR. 2003 Apr;25(4):313-23
references.pmid: 12766393
references.citation: Rev Sci Instrum. 2005 Sep;76(9):1-7
references.pmid: 16508692
references.citation: Biochemistry. 2006 Mar 7;45(9):2820-35
references.pmid: 16503637
references.citation: Gene. 2007 Aug 15;398(1-2):234-48
references.pmid: 17570619
references.citation: Biochemistry. 2000 Dec 26;39(51):16099-109
references.pmid: 11123938
references.citation: J Am Chem Soc. 2005 Jul 27;127(29):10176-7
references.pmid: 16028922
references.citation: Proc Natl Acad Sci U S A. 1990 Jan;87(1):205-9
references.pmid: 2296580
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 10;99(25):16047-51
references.pmid: 12444262
references.citation: Biochim Biophys Acta. 2010 Feb;1798(2):150-60
references.pmid: 19665988
references.citation: J Phys Chem B. 2009 Jun 25;113(25):8560-8
references.pmid: 19485407
references.citation: J Colloid Interface Sci. 2005 Jul 1;287(1):312-7
references.pmid: 15914179
references.citation: Biochemistry. 1985 Oct 8;24(21):5907-18
references.pmid: 4084499
references.citation: Biochim Biophys Acta. 2010 Jan;1804(1):3-14
references.pmid: 19577666
references.citation: Biochemistry. 2008 Jan 15;47(2):517-25
references.pmid: 18085800
references.citation: J Am Chem Soc. 2007 Nov 21;129(46):14311-8
references.pmid: 17958424
references.citation: Biophys Chem. 1996 Jun 11;60(3):63-77
references.pmid: 8679927
references.citation: Eur J Biochem. 1982 Jun 15;125(1):175-81
references.pmid: 6286308
references.citation: Biochemistry. 2012 Jul 31;51(30):5932-41
references.pmid: 22769585
references.citation: Biol Chem. 2008 Sep;389(9):1201-7
references.pmid: 18713007
references.citation: Biophys J. 2010 Apr 7;98(7):1321-6
references.pmid: 20371332
references.citation: Nat Struct Mol Biol. 2011 Feb;18(2):245-9
references.pmid: 21196937
references.citation: Colloids Surf B Biointerfaces. 2011 Mar;83(1):116-21
references.pmid: 21131182
references.citation: Biopolymers. 1999 Aug;50(2):153-61
references.pmid: 10380339
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 17;103(42):15469-72
references.pmid: 17030792
references.citation: J Phys Chem B. 2006 Jun 22;110(24):11615-9
references.pmid: 16800453
references.citation: J Inorg Biochem. 2005 Jan;99(1):72-96
references.pmid: 15598493