title: Organization and dynamics of tryptophan residues in brain spectrin: novel insight into conformational flexibility.
abstract: Brain spectrin enjoys overall structural and sequence similarity with erythroid spectrin, but less is known about its function. We utilized the fluorescence properties of tryptophan residues to monitor their organization and dynamics in brain spectrin. Keeping in mind the functional relevance of hydrophobic binding sites in brain spectrin, we monitored the organization and dynamics of brain spectrin bound to PRODAN. Results from red edge excitation shift (REES) indicate that the organization of tryptophans in brain spectrin is maintained to a considerable extent even after denaturation. These results are supported by acrylamide quenching experiments. To the best of our knowledge, these results constitute the first report of the presence of residual structure in urea-denatured brain spectrin. We further show from REES and time-resolved emission spectra that PRODAN bound to brain spectrin is characterized by motional restriction. These results provide useful information on the differences between erythroid spectrin and brain spectrin.
authors.affiliation: Biophysics and Structural Genomics Division, Saha Institute of Nuclear Physics, 1/AF Bidhannagar, Kolkata, 700064, India.
authors.forename: Madhurima
authors.initials: M
authors.lastname: Mitra
authors.forename: Arunima
authors.initials: A
authors.lastname: Chaudhuri
authors.forename: Malay
authors.initials: M
authors.lastname: Patra
authors.forename: Chaitali
authors.initials: C
authors.lastname: Mukhopadhyay
authors.forename: Abhijit
authors.initials: A
authors.lastname: Chakrabarti
authors.forename: Amitabha
authors.initials: A
authors.lastname: Chattopadhyay
mesh_terms: D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D002942:Circular Dichroism; D005453:Fluorescence; D057927:Hydrophobic and Hydrophilic Interactions; D011487:Protein Conformation; D011489:Protein Denaturation; D012756:Sheep; D013049:Spectrin / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods; D014364:Tryptophan / Q000737:chemistry*; D014508:Urea / Q000737:chemistry
chemical_list: D013049:Spectrin; C038517:prodan; D014364:Tryptophan; D014508:Urea; D015081:2-Naphthylamine
references.citation: J Biol Chem. 2004 Dec 31;279(53):55080-8
references.pmid: 15492010
references.citation: Biochem Biophys Res Commun. 1996 Sep 13;226(2):495-7
references.pmid: 8806662
references.citation: J Biol Chem. 2010 May 7;285(19):14572-84
references.pmid: 20228407
references.citation: Eur Biophys J. 2010 Sep;39(10):1421-32
references.pmid: 20306315
references.citation: Cell Mol Biol Lett. 2001;6(3):691-702
references.pmid: 11598642
references.citation: Methods Biochem Anal. 1991;35:127-205
references.pmid: 2002770
references.citation: J Biol Chem. 1990 Jul 15;265(20):11827-32
references.pmid: 2195026
references.citation: J Biol Chem. 2006 Apr 14;281(15):10527-32
references.pmid: 16476728
references.citation: Physiol Rev. 2001 Jul;81(3):1353-92
references.pmid: 11427698
references.citation: Biochemistry. 1996 Oct 15;35(41):13426-33
references.pmid: 8873611
references.citation: PLoS One. 2013 Dec 20;8(12):e83752
references.pmid: 24376740
references.citation: Acc Chem Res. 2014 Jan 21;47(1):12-9
references.pmid: 23981188
references.citation: Biochemistry. 1995 Oct 10;34(40):13252-8
references.pmid: 7548089
references.citation: Biochim Biophys Acta. 1987 Nov 2;904(1):55-60
references.pmid: 3663667
references.citation: Biochemistry. 1971 Aug 17;10(17):3254-63
references.pmid: 5119250
references.citation: Protein Sci. 2011 May;20(5):867-79
references.pmid: 21412925
references.citation: Proc Natl Acad Sci U S A. 1970 Apr;65(4):823-30
references.pmid: 16591825
references.citation: Protein J. 2004 Jan;23(1):79-83
references.pmid: 15115185
references.citation: Biochem J. 1999 Feb 15;338 ( Pt 1):83-90
references.pmid: 9931302
references.citation: Biochem Biophys Res Commun. 2013 May 24;435(1):64-8
references.pmid: 23618861
references.citation: J Biol Chem. 1997 Aug 22;272(34):21052-9
references.pmid: 9261107
references.citation: Cell Motil Cytoskeleton. 2003 Jan;54(1):16-28
references.pmid: 12451592
references.citation: Science. 2001 Jul 20;293(5529):487-9
references.pmid: 11463915
references.citation: Eur Biophys J. 2010 Sep;39(10):1453-63
references.pmid: 20372885
references.citation: Mol Cell Biol. 1988 Jan;8(1):1-9
references.pmid: 3336352
references.citation: Methods Enzymol. 2008;450:59-78
references.pmid: 19152856
references.citation: Biochemistry. 2006 Nov 14;45(45):13670-6
references.pmid: 17087521
references.citation: Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1299-303
references.pmid: 8108405
references.citation: Biochemistry. 1997 Jul 22;36(29):8977-91
references.pmid: 9220986
references.citation: Biochem Biophys Res Commun. 2010 Apr 16;394(4):1082-6
references.pmid: 20346348
references.citation: Cell Motil Cytoskeleton. 1994;29(1):72-81
references.pmid: 7820859
references.citation: Biopolymers. 2005 Apr 15;77(6):325-34
references.pmid: 15648086
references.citation: Biophys J. 2004 Aug;87(2):831-43
references.pmid: 15298892
references.citation: J Mol Biol. 1997 Oct 31;273(3):740-51
references.pmid: 9356261
references.citation: Blood. 1993 Jun 15;81(12):3173-85
references.pmid: 8507859
references.citation: J Biomol Struct Dyn. 2014;32(6):852-65
references.pmid: 24404769
references.citation: FEBS Lett. 1983 Mar 7;153(1):165-8
references.pmid: 6825856
references.citation: Eur Biophys J. 1988;16(2):121-9
references.pmid: 3208709
references.citation: FEBS Lett. 1999 Nov 5;460(3):391-4
references.pmid: 10556504
references.citation: Cell. 1982 Oct;30(3):725-33
references.pmid: 7139713
references.citation: Biochem J. 1992 Feb 15;282 ( Pt 1):75-80
references.pmid: 1540147
references.citation: Chem Phys Lipids. 2003 Jan;122(1-2):3-17
references.pmid: 12598034
references.citation: Biochemistry. 1997 Nov 25;36(47):14291-305
references.pmid: 9398147
references.citation: Biochemistry. 1997 Jun 10;36(23):6977-85
references.pmid: 9188694
references.citation: Annu Rev Cell Biol. 1993;9:27-66
references.pmid: 8280463
references.citation: J Phys Chem B. 2008 Nov 6;112(44):14075-82
references.pmid: 18842019
references.citation: J Biol Chem. 1990 Mar 15;265(8):4434-43
references.pmid: 1689726
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Crit Rev Biochem Mol Biol. 2005 Jul-Aug;40(4):181-9
references.pmid: 16126485
references.citation: PLoS One. 2015 Jan 24;10(1):e0116991
references.pmid: 25617632
references.citation: J Fluoresc. 1995 Sep;5(3):237-46
references.pmid: 24226791
references.citation: Biochim Biophys Acta. 2002 Aug 31;1564(2):403-11
references.pmid: 12175923
references.citation: J Cell Biol. 2006 Nov 6;175(3):491-503
references.pmid: 17088429
references.citation: Biochemistry. 2003 Dec 16;42(49):14702-10
references.pmid: 14661984
references.citation: J Phys Chem B. 2011 May 19;115(19):5693-706
references.pmid: 21428321
references.citation: Biochemistry. 2009 Jan 13;48(1):206-15
references.pmid: 19072330
references.citation: Protein Sci. 2003 Nov;12(11):2389-403
references.pmid: 14573853
references.citation: Curr Biol. 2007 Mar 20;17(6):562-8
references.pmid: 17331725
references.citation: Science. 2002 Mar 1;295(5560):1719-22
references.pmid: 11872841
references.citation: Biophys J. 1999 Mar;76(3):1469-79
references.pmid: 10049328
references.citation: Biosci Rep. 2006 Dec;26(6):369-86
references.pmid: 17029004
references.citation: Biochemistry. 1979 Jul 10;18(14):3075-8
references.pmid: 465454