Study on the Electrochemical Behaviors of Vincristine and the Interaction of Vincristine with Tubulin

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1 , ACTA CHIMICA SINICA Vol 62, 2004 No 2, a, b Ξ, a a a Ξ ( a ) ( b ) 0105 mol/ L Tris, 0115 mol/ L NaCl, (VCR) V (vs Ag/ AgCl), mol/ L VCR, mol/ L, VCR Frumkin VCR,VCR,,,, Study on the Electrochemical Behaviors of Vincristine and the Interaction of Vincristine with Tubulin YU, Yong a, b HU, Jing2Bo Ξ, a SHANG, Jun a LI, Qi2Long a ( a Department of Chemistry, Beijing Normal University, Beijing ) ( b National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, Beijing ) Abstract A sensitive reduction peak of vincristine (VCR) is obtained by adsorptive stripping voltammetry in 0105 mol/ L Tris and 0115 mol/ L NaCl buffer solution The peak potential is V (vs Ag/ AgCl) The peak current is directly proportional to the concentration of VCR ( mol/ L), with a detection limit of mol/ L The electrochemical behavior of this system was studied by the methods of normal pulse polarography, linear2sweep and cyclic voltammetry The reduction process is irreversible with adsorptive characteristics The adsorption of VCR obeys the Frumkin isotherm The electrochemical behavior of the binding of VCR to tubulin was studied by cyclic voltammetry The results showed that the reaction of tubulin dimer with VCR formed an electrochemically active complex, and the reduction process of the complex is irreversible Keywords vincristine (VCR), voltammetry, adsorption, tubulin (Vincristine, VCR) VCR 11,, 111 HPLC, [1] ( EG&G Princeten ), VCR : [2],,,Ag/ AgCl, [3],HPLC [1] VCR [4], VCR [5] 1 Ξ E2mail : edu cn ; Tel : (010) ; Fax : (010) Received March 23, 2003 ; revised July 14, 2003 ; accepted September 12, 2003 (No )

2 138 Vol 62, VCR Figure 1 Structure of VCR nm, JP303 ( ),, - Beckman DU640 Beckman L82M Ultracen trifuge Beckman DU640, Bradford, 807 ph/ ( 16 5 mg/ ml,, 2, ) 501 ( ) [6], 95 %, c tubulin = mol/ L VCR mol/ L, mol/ L Tris (ph = 7 20) 115 mol/ L NaCl (MES) (100 mmol/ L MES, 1 mmol/ L EGTA, 015 mmol/ L MgCl 2, 1 mmol/ L GTP ATP) 2MES ( 100 mmol/ L MES, 1 mmol/ L EGTA, 015 mmol/ L MgCl 2, 8 mol/ L,1 mmol/ L [6 ] mol/ L Tris (ph = 7 20), 0115 mol/ L NaCl, 4, V 300 s, 100 mv/ s,,, VCR mol/ L, cm 2, MES,500 r/ min 1 2 ; ml MES , g 1 h,, SDS, SDS, SDS, 10 %,, 95 % Bradford [8] :, ( max ) nm, mol/ L Tris 0115 mol/ L NaCl,, 0105 mol/ L Tris (ph = 7 20) 0115 mol/ L NaCl, VCR GTP ATP), 1 mol/ L NaOH ph 6 5, GTP ATP 1 mmol/ L V (vs Ag/ AgCl), ; V,, V, V ( R = ), mol/ L 2 3 t < 44, t > 2MES, min( 44,, ), 26, g 1 h,, ;,, 1/ 10 MES,, t < 44, 1 h, ;, ; t > 44, -, %/, 35, MES, %/, %/, mol/ L VCR V s, 100 mv/ s, SDS [7] :, SDS, VCR ;, VCR

3 No 2 [9] : 139 = s - 1 n = 0185, 0 < < 015, n = 2, = 01425, n = 0185, E p ln v 2 Figure 2 Repetitive cyclic voltammograms 0105 mol/ L Tris (ph = 7 20), 0115 mol/ L NaCl, E i = V (vs Ag/ AgCl), v = 100 mv/ s, mol/ L VCR VCR 2 5 : V,, VCR [10] VCR Q, = Q/ ( nfa), A I pc t acc, ( mol/ L VCR 180 s, , VCR Q mol/ L VCR VCR, s), VCR [9] 10-7 mol/ L, VCR = mol/ cm 2 VCR, 2 3 5, I pc, c VCR = mol/ L, I pc v, v 1/ 2, VCR, ; c VCR = mol/l, I pc v, v 1/ 2, 0 2, VCR VCR c I p / c VCR, 25, G 0 = kj/ mol, 2 6 VCR 2 4 n k mol/ L Tris (ph = 7 20), mol/ L NaCl 4 VCR VCR2,, E p v 4A mol/ L Tris, 0105 mol/ L NaCl (ph [11] : E p = E 01 + RT nf ln RT k0 - nf ln v 3 ln v Figure 3 Plot of the peak potential vs ln v mol/ L Tris (ph = 7 20), mol/ L NaCl, mol/ L VCR, other conditions are same as in Figure 2, VCR / cm c(1 - - ln ), VCR Frumkin = 1118, = L/ mol = exp ( - G 0 / RT) 7110), c VCR = mol/ L VCR,, V (vs Ag/ AgCl) mol/ L, 4B,, V, VCR, E p ln v ( 3), 4B,, RT/ nf = E p v E 01 = ;,, VCR,VCR V, = V,

4 140 Vol 62, (TNB), 1 TNB (DTT) [12] : VCR2, 4 VCR VCR2tubulin Figure 4 Repetitive cyclic voltammograms of VCR and VCR2tubulin VCR dimer complex VCR A mol/ L Tris, 0105 mol/ L NaCl (ph = 7 10), c VCR = mol/ 5 6 VCR L, v = 100 mv/ s, E i = (vs Ag/ AgCl) at 37, t acc = 100 s ; B A + c tubulin = mol/ L DTT DTNB, DTT DTNB, VCR, c VCR = mol/ L, c tubulin = mol/ L, t acc,vcr2 I pc t acc, t acc = 500 s,, VCR I pc, c VCR = mol/ L, c tubulin = mol/ L, I pc v, v 1/ 2,, ; c VCR = mol/ L, c tubulin = mol/ L, I pc v, v 1/ 2,, VCR2, VCR 5, Figure 5 Absorption spectra at 37 s = mol/ L Tris and 0105 mol/ L NaCl, ph = 7 10 ; (a) s ,5-222 (DTNB), Ellman mol/ L VCR vs s ; (b) s , - 6 mol/ L tubulin vs s + MES DTNB, - 6 mol/ L tubulin vs s + MES 85 L L ; (c) a

5 No 2 : References 6 Figure 6 Absorption spectra at 37 s = mol/ L Tris and 0105 mol/ L NaCl (ph = 7 10) ; (a) s mol/ L VCR mol/ L tubulin vs s mol/ L tubulin ; (b) s mol/ L VCR mol/ L tubulin vs s mol/ LVCR + MES 85 L 1 Castle, M C ; Mead, J A Biochem Pharmacol 1978, 27, 5, 37,01005 mol/ L Tris, 0105 mol/ L NaCl (ph = 7 10),VCR - 8 Ausubel, F M ; Brent, R ; Kingston, R E Short Protocols In Molecular Biology, 3rd ed, John Wiley & Sons, New York, 190, 206, 220, nm, 1995, p 10 9 Li, Q L ; Chen, S A Anal 310, nm VCR Chim Acta 1993, 282, Anson, F ( translated by Huang, W Z ), Electrochemistry and Electroanalytical Chemistry, Peking University Press, Beijing, nm, VCR + tubulin < VCR , p 45 (in Chinese) tubulin, 5 a 6 a (Anson, F,,, VCR - ; 5 b 6 b,, 1983, p 45 ) -, 11 Ji, G ; Li, Q L Chem J Chin Univ 1991, 11, 1642 (in, Chinese) a a, b b a a, 6 (,,, 1991, 11, 1642 ) a 210 nm - 12 Zhou, H M ; Wang, H R Modification of Proteins, Tsinghua, University Press, Beijing, 1998, p 1, p 19 (in Chinese) ;b b, (,,,,,, 1998, p 1, p 19 ) 37 2 Carmel, R Am J Clin Pathol 1978, 69, Dixon, G J ; Dulmadge, E A ; Mellet, L B Am J Clin Pathal 1969, 29, Temizer, A Talanta 1986, 32 (10), Wang, J ; Zadeii, J ; Shanlin, M J Electroanal Chem 1987, 237, Li, Z R ; Liu, Z L ; Sun, Y H ; Han, R ; Yin, M B ; Ye, Q R Acta Pharmaceutica Sinica 1986, 21, 651 ( in Chinese) (,,,,,,, 1986, 21, 651 ) 7 Sambrook, J ; Fritsch, E F ; Maniatis, T (translated by Jin, D Y ; Li, M F ) Molecular Cloning : A Laboratory Manual 2nd ed, Science Press, Beijing, 1995, p 880 (in Chinese) ( J, E F, T,,, ( ),,, 1995, p 880 ) (A LU, Y J ; FAN, Y Y )

6 Graphical Abstract Vol 62, 2004 Synthesis of Pt Nano2particles Modified with Cation Ions and Its Self2assembly on Nafion Membrane TANG, Hao2Lin ; PAN, Mu ; YUAN, Run2Zhang Acta Chimica Sinica 2004, 62 (2), 127 Pt particles modified with quaternary ammonium cation were electrostatic self2assembled on the surface of Nafion membrane A reasonable mathematic model was put forward and the results were used to explain the experiment results Application in Image Processing of Nonlinear Light2excitable Membrane Containing Ru2 ( bpy) 3 Cl 2 GU, Qiang ; ZOU, Ai2Hua ; WANG, Yong ; YUAN, Chun2Wei Acta Chimica Sinica 2004, 62 (2), 131 The light2excitable membrane containing Ru ( bpy) 3 Cl 2 as photocatalyst could catalyze the Belousov2Zhabotinsky ( BZ) reaction by illumination A fine2definition image processing could be auto2performed when projecting a picture on this medium The boundary of the image formed in membrane was enhanced evidently Study on the Electrochemical Behaviors of Vincristine and the Interaction of Vincristine with Tubulin YU, Yong ; HU, Jing2Bo ; SHANG, Jun ; LI, Qi2 Long Acta Chimica Sinica 2004, 62 (2), 137 A sensitive reduction peak of vincristine (VCR) is obtained by adsorptive stripping voltammetry in 0105 mol/ L Tris and 0115 mol/ L NaCl buffer solution The peak potential is V (vs Ag/ AgCl) The peak current is directly proportional to the concentration of VCR, with a detection limit of mol/ L The electrochemical behavior of this system was studied by the methods of normal pulse polarography, linear2sweep and cyclic voltammetry Photodegradation Dynamics of Pure Micro2 cystin Variants with Illumination of Fixed Wavelength UV2lights CHEN, Wei ; GAN, Nan2Qin ; SONG, Li2Rong Acta Chimica Sinica 2004, 62 (2), 142 Chromatograms of MC2LR and MC2RR at different time after lighting with weaker UV irradiation