2004 62 2, 137 141 ACTA CHIMICA SINICA Vol 62, 2004 No 2, 137 141 a, b Ξ, a a a Ξ ( a 100875) ( b 100080) 0105 mol/ L Tris, 0115 mol/ L NaCl, (VCR) - 1168 V (vs Ag/ AgCl), 110 10-8 210 10-7 mol/ L VCR, 710 10-9 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 100875) ( b National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, Beijing 100080) 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 - 1168 V (vs Ag/ AgCl) The peak current is directly proportional to the concentration of VCR (110 10-8 210 10-7 mol/ L), with a detection limit of 710 10-9 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] 37028 ( EG&G Princeten ), VCR : [2],,,Ag/ AgCl, [3],HPLC [1] VCR [4], VCR [5] 1 Ξ E2mail : hujingbo @bnu edu cn ; Tel : (010) 62209398 ; Fax : (010) 62200567 Received March 23, 2003 ; revised July 14, 2003 ; accepted September 12, 2003 (No 20275007)
138 Vol 62, 2004 1 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 ( ) 112000 [6], 95 %, c tubulin = 1140 112 10-4 mol/ L VCR 110 10-3 mol/ L, 2 2 110 mol/ L Tris (ph = 7 20) 115 mol/ L NaCl 22 - - (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 2 2 1 2 2 3 [6 ] 2 1 1 0105 mol/ L Tris (ph = 7 20), 0115 mol/ L NaCl, 4, - 1120 V 300 s, 100 mv/ s,,, VCR 110 10-8 210 10-7 mol/ L, 015 1 cm 2, MES,500 r/ min 1 2 ; 015 1 ml MES 4 2 3 1 4, 105000 g 1 h,, SDS, SDS, SDS, 10 %,, 95 % 2 1 3 Bradford [8] :, ( max ) 455 595 nm, 595 2 2 1 0105 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, 2 2 2 GTP ATP 1 mmol/ L - 111 V (vs Ag/ AgCl), ; - 111-113 V,, - 113 V, - 112 V ( R = 019969), 710 10-9 mol/ L 2 3 t < 44, t > 2MES, 37 30 min( 44,, ), 26, 105000 g 1 h,, ;,, 1/ 10 MES,, t < 44, 1 h, ;, ; t > 44, -, 9 35 01063 %/, 35, MES, 44 01024 %/, 44 50-01015 %/, 2 3 2 2 9 87 10-8 mol/ L VCR - 1120 V 2 1 2 300 s, 100 mv/ s, SDS [7] :, SDS, VCR ;, VCR
No 2 [9] : 139 = 01010 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 = - 1120 V (vs Ag/ AgCl), v = 100 mv/ s, 9 87 10-8 mol/ L VCR 2 3 3 VCR 2 5 : - 1164 V,, VCR [10] 2 5 1 VCR 2 3 4 Q, = Q/ ( nfa), A I pc t acc, (9 87 10-8 mol/ L VCR 180 s, 1141 10-6, VCR Q mol/ L VCR VCR, 2 26 60 s), VCR [9] 10-7 mol/ L, VCR = 9 75 10-8 mol/ cm 2 VCR, 2 3 5, I pc, c VCR = 9 87 10-8 mol/ L, I pc v, v 1/ 2, VCR, ; c VCR = 1141 10-6 mol/l, I pc v, v 1/ 2, 0 2, VCR 2 3 6 VCR c I p / c VCR, 25, G 0 = - 37 47 kj/ mol, 2 6 VCR 2 4 n k 0 2 6 1 01005 mol/ L Tris (ph = 7 20), 01015 mol/ L NaCl 4 VCR VCR2,, E p v 4A 01005 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 01005 mol/ L Tris (ph = 7 20), 01015 mol/ L NaCl, 1112 10-7 mol/ L VCR, other conditions are same as in Figure 2, VCR 117 10-17 / cm 2 2 5 2 c(1 - - ln ), VCR Frumkin = 1118, = 3 7 10 6 L/ mol = exp ( - G 0 / RT) 7110), c VCR = 2106 10-6 mol/ L VCR,, - 1155 V (vs Ag/ AgCl) 1140 10-6 mol/ L, 4B,, - 1167 V, VCR, E p ln v ( 3), 4B,, RT/ nf = 01033 E p v E 01 = ;,, VCR,VCR2-11525 V, = - 1176 V,
140 Vol 62, 2004 1 22 252 (TNB), 1 TNB (DTT) [12] : VCR2, 4 VCR VCR2tubulin Figure 4 Repetitive cyclic voltammograms of VCR and VCR2tubulin VCR dimer complex 2 6 5 VCR A 01005 mol/ L Tris, 0105 mol/ L NaCl (ph = 7 10), c VCR = 2106 10-6 mol/ 5 6 VCR L, v = 100 mv/ s, E i = - 1130 (vs Ag/ AgCl) at 37, t acc = 100 s ; B A + c tubulin = 1140 10-6 mol/ L DTT DTNB, DTT DTNB, VCR, 2 6 2 c VCR = 2 68 10-7 mol/ L, c tubulin = 1140 10-7 mol/ L, t acc,vcr2 I pc t acc, t acc = 500 s,, VCR 2 6 3 I pc, c VCR = 268 10-7 mol/ L, c tubulin = 1140 10-7 mol/ L, I pc v, v 1/ 2,, ; c VCR = 2106 10-6 mol/ L, c tubulin = 1140 10-6 mol/ L, I pc v, v 1/ 2,, 2 6 4 VCR2, VCR 5, Figure 5 Absorption spectra at 37 s = 01005 mol/ L Tris and 0105 mol/ L NaCl, ph = 7 10 ; (a) s + 2 68 10-6 5,5-222 (DTNB), Ellman mol/ L VCR vs s ; (b) s + 1120 10, - 6 mol/ L tubulin vs s + MES 85 + 1120 10 DTNB, - 6 mol/ L tubulin vs s + MES 85 L L ; (c) a
No 2 : 141 - References 6 Figure 6 Absorption spectra at 37 s = 01005 mol/ L Tris and 0105 mol/ L NaCl (ph = 7 10) ; (a) s + 2 68 10-6 mol/ L VCR + 1120 10-6 mol/ L tubulin vs s + 1120 10-6 mol/ L tubulin ; (b) s + 2 68 10-6 mol/ L VCR + 1120 10-6 mol/ L tubulin vs s + 2 68 10-6 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, 253 298 nm, 1995, p 10 9 Li, Q L ; Chen, S A Anal 310, 280 194 nm VCR Chim Acta 1993, 282, 145 10 Anson, F ( translated by Huang, W Z ), Electrochemistry and Electroanalytical Chemistry, Peking University Press, Beijing, 190 300 nm, VCR + tubulin < VCR + 1983, 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, 137 3 Dixon, G J ; Dulmadge, E A ; Mellet, L B Am J Clin Pathal 1969, 29, 1810 4 Temizer, A Talanta 1986, 32 (10), 792 5 Wang, J ; Zadeii, J ; Shanlin, M J Electroanal Chem 1987, 237, 2811 6 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 ) (A0303208 LU, Y J ; FAN, Y Y )
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 - 1168 V (vs Ag/ AgCl) The peak current is directly proportional to the concentration of VCR, with a detection limit of 710 10-9 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