title: Mechanistic investigation of domain specific unfolding of human serum albumin and the effect of sucrose.
abstract: This study is devoted to understand the unfolding mechanism of a multidomain protein, human serum albumin (HSA), in absence and presence of the sucrose by steady-state and time-resolved fluorescence spectroscopy with domain specific marker molecules and is further being substantiated by molecular dynamics (MD) simulation. In water, the domain III of HSA found to unfold first followed by domains I and II as the concentration of GnHCl is increased in the medium. The sequential unfolding behavior of different domains of HSA remains same in presence of sucrose; however, a higher GnHCl concentration is required for unfolding, suggesting stabilizing effect of sucrose on HSA. Domain I is found to be most stabilized by sucrose. The stabilization of domain II is somewhat similar to domain I, but the effect of sucrose on domain III is found to be very small. MD simulation also predicted a similar behavior of sucrose on HSA. The stabilizing effect of sucrose is explained in terms of the entrapment of water molecules in between HSA surface and sucrose layer as well as direct interaction between HSA and sucrose.
authors.affiliation: Department of Chemistry, Indian Institute of Technology Kanpur, Kanpur, 208 016, Uttar Pradesh, India.
authors.forename: Rajeev
authors.initials: R
authors.lastname: Yadav
authors.forename: Pratik
authors.initials: P
authors.lastname: Sen
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D000818:Animals; D002942:Circular Dichroism; D019791:Guanidine / Q000737:chemistry; D006427:Hemin / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D011489:Protein Denaturation; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding; D012709:Serum Albumin / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013395:Sucrose / Q000378:metabolism*; D014364:Tryptophan / Q000737:chemistry; D014867:Water
chemical_list: C515594:8-anilino-1-naphthalenesulfonic acid; D000817:Anilino Naphthalenesulfonates; D012709:Serum Albumin; D014867:Water; D013395:Sucrose; D006427:Hemin; D014364:Tryptophan; D019791:Guanidine
keywords: denaturation; human serum albumin; multidomain protein; stabilization; sugar; unfolding mechanism
references.citation: J Phys Chem B. 2008 Apr 10;112(14):4372-80
references.pmid: 18341329
references.citation: Eur J Biochem. 1997 Aug 15;248(1):24-9
references.pmid: 9310355
references.citation: Protein Sci. 2011 Dec;20(12):2095-101
references.pmid: 21997768
references.citation: J Biol Chem. 2001 Jun 22;276(25):22804-9
references.pmid: 11285262
references.citation: J Phys Chem B. 2007 Apr 5;111(13):3563-9
references.pmid: 17388507
references.citation: Pharm Res. 1991 Jul;8(7):888-92
references.pmid: 1924140
references.citation: Protein Sci. 2013 Apr;22(4):467-74
references.pmid: 23389799
references.citation: Biol Pharm Bull. 2002 Jun;25(6):695-704
references.pmid: 12081132
references.citation: Chemphyschem. 2009 Jul 13;10(9-10):1664-71
references.pmid: 19466702
references.citation: J Comput Chem. 2005 Dec;26(16):1668-88
references.pmid: 16200636
references.citation: J Comput Chem. 2004 Jul 15;25(9):1157-74
references.pmid: 15116359
references.citation: Z Naturforsch C J Biosci. 1994 May-Jun;49(5-6):327-30
references.pmid: 8060458
references.citation: FEBS Lett. 1997 May 12;408(1):67-70
references.pmid: 9180270
references.citation: J Biol Chem. 1999 Oct 8;274(41):29303-10
references.pmid: 10506189
references.citation: Eur J Biochem. 2004 May;271(9):1789-97
references.pmid: 15096218
references.citation: J Fluoresc. 1996 Mar;6(1):33-40
references.pmid: 24226995
references.citation: J Biochem. 1987 Aug;102(2):313-7
references.pmid: 3667572
references.citation: Eur J Biochem. 1994 Jan 15;219(1-2):187-93
references.pmid: 8306985
references.citation: Protein Eng. 1999 Jun;12(6):439-46
references.pmid: 10388840
references.citation: J Protein Chem. 1990 Aug;9(4):461-5
references.pmid: 2275755
references.citation: Proc Natl Acad Sci U S A. 1996 Dec 10;93(25):14411-5
references.pmid: 8962065
references.citation: J Biol Chem. 1973 Apr 25;248(8):2650-5
references.pmid: 4697387
references.citation: J Comput Chem. 2009 Oct;30(13):2157-64
references.pmid: 19229944
references.citation: Int J Biol Macromol. 2008 Jun 1;42(5):483-90
references.pmid: 18452986
references.citation: Proc Natl Acad Sci U S A. 2004 Sep 14;101(37):13411-6
references.pmid: 15353599
references.citation: Mol Pharmacol. 1988 Aug;34(2):160-71
references.pmid: 3412320
references.citation: Protein Sci. 2004 Nov;13(11):2949-59
references.pmid: 15459332
references.citation: Biochim Biophys Acta. 2007 Jul;1774(7):928-35
references.pmid: 17581805
references.citation: Biophys Chem. 2009 Sep;144(1-2):46-52
references.pmid: 19573978
references.citation: Biochemistry. 1989 May 2;28(9):3916-22
references.pmid: 2526652
references.citation: Mol Cell Biochem. 2004 Jan-Feb;256-257(1-2):319-27
references.pmid: 14977191
references.citation: Adv Protein Chem. 2003;66:27-85
references.pmid: 14631816
references.citation: Proteins. 2000 Jul 1;40(1):29-38
references.pmid: 10813828
references.citation: Pharm Res. 1997 Nov;14(11):1607-12
references.pmid: 9434282
references.citation: Eur J Biochem. 2003 Jun;270(11):2497-504
references.pmid: 12755705
references.citation: Mol Pharmacol. 1976 Nov;12(6):1052-61
references.pmid: 1004490
references.citation: J Am Chem Soc. 2008 Aug 13;130(32):10793-801
references.pmid: 18642807
references.citation: Int J Biol Macromol. 2005 Dec 15;37(4):200-4
references.pmid: 16324740
references.citation: Arch Biochem Biophys. 1999 May 15;365(2):289-98
references.pmid: 10328824
references.citation: BMC Struct Biol. 2003 Jul 7;3:6
references.pmid: 12846933
references.citation: Biopolymers. 1994 Jul;34(7):957-61
references.pmid: 8054473
references.citation: Anal Sci. 2009 Jan;25(1):115-20
references.pmid: 19139584
references.citation: Arch Biochem Biophys. 1998 Dec 1;360(1):10-4
references.pmid: 9826423
references.citation: Proc Natl Acad Sci U S A. 1982 Jan;79(1):71-5
references.pmid: 6275391
references.citation: Protein Sci. 1993 Sep;2(9):1490-6
references.pmid: 8104614
references.citation: J Pharm Sci. 1996 Oct;85(10):1131-2
references.pmid: 8897285
references.citation: Biomacromolecules. 2010 Jan 11;11(1):106-12
references.pmid: 19899798
references.citation: Biophys Chem. 1992 Nov;45(1):1-6
references.pmid: 1467440
references.citation: Biophys Chem. 1996 Jun 11;60(3):63-77
references.pmid: 8679927
references.citation: Biochim Biophys Acta. 2005 Jun 15;1750(1):93-102
references.pmid: 15890566
references.citation: Biomacromolecules. 2009 Mar 9;10(3):517-21
references.pmid: 19173654
references.citation: Biochim Biophys Acta. 2004 Aug 4;1673(3):139-48
references.pmid: 15279885
references.citation: Phys Chem Chem Phys. 2011 Feb 14;13(6):2294-9
references.pmid: 21116551
references.citation: Nat Struct Biol. 1998 Sep;5(9):827-35
references.pmid: 9731778
references.citation: Nature. 1992 Jul 16;358(6383):209-15
references.pmid: 1630489
references.citation: Chemphyschem. 2003 Aug 18;4(8):824-9
references.pmid: 12961979
references.citation: Adv Protein Chem. 1968;23:121-282
references.pmid: 4882248
references.citation: Proteins. 2004 Apr 1;55(1):177-86
references.pmid: 14997551
references.citation: Biochemistry. 1988 Oct 18;27(21):8063-8
references.pmid: 3233195