CHEM ICAL JOU RNAL O F CH IN ESE UN IV ERS IT IES [1, 2 ]. ., FT IR, , g ( cm - 1 ). H 2O cm - 1. :

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1 V ol. 24 N o CHEM ICAL JOU RNAL O F CH IN ESE UN IV ERS IT IES , (, ) BSA R ase A, : cm - 1 Α2 ; cm - 1 Β2 ; cm - 1 ; cm - 1 Β2.,, X. (FT IR ); ; ; O A (2003) [1, 2 ].,. X ; ; ; ( T rp, T yr ) [3, 4 ] ;,, [5 ]., [6 ]., (FT IR ) [7, 8 ].., FT IR,,., ( cm - 1 ). H 2O cm - 1. [9 ],.,, H 2O,.,.,.. : Byler [10 ], Surew icz [11, 12 ]. [11 ],,.,,, [12 14 ]., Α2 Β2 [13, 15 ], Β2. Cai Sinh [16 ] cm - 1 Β2, cm - 1, X.,,,, X. : : ( : ). : (1962 ),,,,. E2m ail: m enxia2xie@ 263. net

2 N o. 2 : N exus 670 (N icolet ) ; (A TR, ZnSe ), D T GS, OM N IC 5. 2, Galactic Peak solve. (BSA ) A (R nase A ) A (Concanavalin A ) Si2 m a ; (T ryp sin, G IBCO ) ; C (Cytoch rom e C, ) ; G ( IG, ). N ah 2PO 4 (, ), N aoh (, ), ( F isher, ), M illi2q mmoll N ah 2PO 4, N aoh ph 7. 4,, 2%., N ah 2PO 4 20 mmoll,, 2% , 4 cm , OM N IC 5. 2., A TR ZeSe, 4 cm - 1, 512, , cm - 1 [11 ]. ( cm - 1 ; cm - 1 ), 5 Savitsk2Golay,,., Gausse,.,, ,.,, [17 19 ].,,. Α2 BSA Β2 R nase A. BSA, 20%, 30%, 40% 50%, R nase A 20%, 30% 40%. R nase A Β2, 40%,,,. BSA 50%. BSA 50% R nase A 40% BSA R nase A. 1 3 ( ), 2 cm Α2 BSA, 1 324, 1 316, cm - 1 Α2, 48%, [9, 17 ]., Α2, 0 50%, BSA 48. 4% 38. 3%, ( 2), cm - 1 [ 1 (B) ]. R nase A X 23%,

3 228 V ol. 24, 1 320, cm - 1 Α2, Α2 23%., R nase A Α % 19. 2%, ( 2) cm - 1 [ 2 (B ) ]. Table 1 The inf luence of oran ic solven t on the secondary structure of BSA and Rna se A Peak Position cm - 1 A rea (% ) A ssinm ent of the component peaks secondary structure(% ) BSA Α2Helix Α2Helix Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet BSA Α2Helix % CH 3OH Α2Helix Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet BSA Α2Helix % CH 3OH Α2Helix Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet BSA Α2Helix % CH 3OH Α2Helix Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet BSA Α2Helix % CH 3OH Α2Helix Α2Helix Α2Helix Peak Position cm - 1 A rea (% ) A ssinm ent of the component peaks secondary structure (% ) BSA Β2Turn % CH 3OH Β2Turn Random Random Β2Sheet Β2Sheet Rnase A Β2Sheet Α2Helix Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet Rnase A Α2Helix % CH 3OH Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet Rnase A Α2Helix % CH 3OH Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet Rnase A Α2Helix % CH 3OH Α2Helix Α2Helix Β2Turn Β2Turn Random Random Β2Sheet Β2Sheet Β2Sheet Β2 Β2,.

4 N o. 2 : 229 F i. 1 The curve-f ittin results of am ides (A) and (B) of BSA in 50% CH3OH in volume fraction F i. 2 The curve-f ittin results of am ides (A) and (B) of Rna se A in 40% CH3OH in volume fraction BSA Β %, Β2 1% ; 50%, Β2 4. 6%. 3 Β2 3 cm - 1, cm - 1. R nase A Β2, 40% 11%. 3, cm cm - 1 Β2, cm Β2 Β2,. BSA Β2 2, cm - 1. Table 2 The result of am ide and am ide curve-f ittin Volum e fraction (% ) 3 Band Α2Helix Β2Sheet Β2Turn Random BA S Rnase A Volum e ratio: V CH 3 OHV total., Β2. 0% 50%, 3% cm - 1 ( 1). R nase A,, ( cm - 1 ) Β2, 18. 6%, X 21%., 4%, 2 2 cm - 1., Β cm - 1, cm - 1 Β BSA

5 230 V ol. 24 ( cm - 1 ),,,, 4 cm %, ( 2). R nase A cm - 1, 11. 6%, X 10%., 11. 6% 22. 7%,, 3 cm - 1.,,., cm - 1., : cm - 1 Α2 ; cm - 1 Β2 ; cm - 1 ; cm - 1 Β cm - 1, cm cm - 1 3,.,. Α2,. 2. 2, cm - 1 Α2 ; cm - 1 Β2 ; cm - 1 Β2 ; cm - 1 [8 ]. BSA [17 ] R nase A [20 ] [ 1 (A ) 2 (A ) ],, 2., S : S = [ (x i - y i ) 2 (N - 1) ] 12, x i, y i ; N., S, S,. Α2 Β2 Β2 S R nase A , , , ; BSA , , , ; 0. 03,, X, X. X, 3.,, S. Α2 Β2 Β2, S , , , ; X S , , , , S ,, X,. Table 3 Compar ison of the results obta ined by am ides and and X-ray da ta M ethod BSA Reḟ [ 9, 17 ] RN ase A T ryp sin Concanavalin A IG Cytochrom e C Α2Helix (% ) Β2Sheet (% ) Β2Turn (% ) Random (% )

6 N o. 2 : 231 [ 1 ] YAN G Chi2M in ( ). Chem. J. Chinese U niversities( ) [J ], 2002, 23 (2) : [ 2 ] J IAN G Jun2Guan ( ), WAN G Zhen2Xin ( ), L IU Chan2W ei( ) et al.. Chem. J. Chinese U niversities( ) [J ], 2001, 22 (7) : [ 3 ] L I Xiao2J in ( ), ZHAN G Shan2Ron ( ), ZHAN G Shu2Gon ( ) et al.. Chem. J. Chinese U niversities( ) [J ], 1999, 20 (1) : [ 4 ] ZHAO Chen2Guan ( ), L IU L an2yin ( ), Q IU A i2don ( ) et al.. Chem. J. Chinese U niversities( ) [J ], 1997, 18 (5) : [ 5 ] Parvez İ H., Feride S.. J. M olecular Catalysis B, Enzym atic[j ], 1999, 7: [ 6 ] X IE M en2xia ( ), L IU Yuan ( ). Chinese J. A nalysis L aboratory ( ) [J ], 2001, 20 (Supp l. ) : [ 7 ] Xie M. X.. Spectroscopy L etters[j ], 1996, 29 (1) : [ 8 ] Xie M. X., L iu Y.. Spectrochim ica A cta, Part A [J ], 2002, 58 (13) : [ 9 ] Fu F. N., DeO liveria D. B., T rum ble W. R. et al.. App l. Spectrosc. [J ], 1994, 48: [ 10 ] Byler D. M., B rouillette J. N., Susi H.. Spectroscopy[J ], 1986, 1 (3) : [ 11 ] Surew iczw. K., M oscarello M. A., M antsch H. H.. J. B iochem. [J ], 1987, 262 (18) : [ 12 ] Surew iczw. K., Szabo A., M antsch H. H.. Euṙ J. B iochem. [J ], 1987, 167 (3) : [ 13 ] Don A., Huan P., Cauhey W. S.. B iochem istry[j ], 1990, 29: [ 14 ] Kaiden K., M atsui T., Tanaka S.. Spectroscopy[J ], 1987, 41: [ 15 ] Sinh B. R., Fuller M. P., Schiavo G.. B iophyṡ Chem. [J ], 1990, 36: [ 16 ] Cai S., Sinh B. R.. B iophysical Chem. [J ], 1999, 80: 7 20 [ 17 ] Cyril R.. J. M olecular Structure[J ], 1999, 478: [ 18 ] Jam esm. P., Heino S.. J. B iochem ical and B iophysicalm ethods[j ], 1984, 9: [ 19 ] WAN G B in ( ), WAN G J in ( ), YU J ian ( ) et al.. Spectroscopy and Spectral A nalysis( ) [J ], 1999, 19 (5) : [ 20 ] N eault J. F., Tajim ir2r iahi H. A.. B iochim ica et B iophysica A cta[j ], 1998, 1 384: Stud ies on Infrared Bands for the Secondary Structure D eterm ination of Prote in s X IE M en2x ia 3, L IU Yuan (A naly tical & T estin Center, B eij in N orm al U niversity, B eij in , Ch ina) Abstract Fourier transform infrared spectroscopy is increasinly becom in an important m ethod for quan2 titatively determ inin the secondary structure of p roteinṡ band ( cm - 1 ) and am ide ( cm - 1 ) are two m ain bands for th is purposeṡ w as nelected because of its rela2 tively w eak in sinals, but there is not interference from w ater and w ater vapor vibration bands and it is more sensitive to the chanes of p rotein secondary structure. In th is paper, p roteins BSA and RN ase A w ere denatured by m ethanol. T he component bands of secondary structure in am ide w ere assined by com binin the quantitative results of am ide band. Α2H elix cm - 1 ; Β2turn cm - 1 ; R andom coil cm - 1 and Β2sheet cm - 1. T he overlapp in area betw een the neihborin bands of different structures, such as cm - 1, cm - 1, cm - 1, can be determ ined accordin to quantitative results of am ide band. By usin above assinm ents, the quantitative analysis results of the p roteins in w h ich the secondary structures have been know n w ere consistent w ith that of X2ray data and am ide band. Keywords FT IR; band; Secondary structure of p roteins; Solvent denaturation (Ed. : A, G)