Comparison between HPLC and capillary electrophoresis for the separation and identification of the platination products of oligonucleotides with cis-[Pt(NH3)(2)(H2O)(2)](2+)

被引:23
作者
Troujman, H [1 ]
Chottard, JC [1 ]
机构
[1] UNIV PARIS 05,CHIM & BIOCHIM PHARMACOL & TOXICOL LAB,CNRS,URA 400,F-75270 PARIS 06,FRANCE
关键词
D O I
10.1006/abio.1997.2292
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
HPLC and capillary electrophoresis (CE) were compared in order to separate the platinum adducts formed upon reaction of the double-stranded 18-mer d(AACGGTTAACCGTTAATT)(2) I with the two complexes cis-[Pt(NH3)(2)(H2O)(2)](2+) 1 and [Pt(NH3)(3)(H2O)](2+) 2 and of the single-stranded octamer d(CTGGCTCA) II with complex 2. The GG sequence in both oligonucleotides is the only site of platination giving monoadducts on either of the two guanines or the diadduct which is the N7,N7 chelate of the Pt(NH3)(2)(2+) moiety. The HPLC separation of the adducts was performed with a reverse-phase technique similar to that previously used for platinated oligonucleotides up to octamers. The adducts were identified by enzymatic digestion followed by mass spectrometry characterization of the digests collected at the outlet of the column. The CE separation of the platination products of I with 2 and 1 was performed using respectively a simple capillary zone electrophoresis technique and a micellar electrokinetic capillary chromatography technique. For the more difficult separation of the platination products of II with 2, a capillary gel electrophoresis technique was required. In contrast to HPLC, CE gives better resolution the longer the oligonucleotide. Moreover, the separations are run with higher efficiency and time saving. However, with the present machines, CE precludes the recovery of sufficient amounts of material required by product identification. (C) 1997 Academic Press.
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页码:177 / 185
页数:9
相关论文
共 29 条
[21]   CALIBRATION OF POLYACRYLAMIDE-GEL COLUMNS FOR THE SEPARATION OF OLIGONUCLEOTIDES BY CAPILLARY ELECTROPHORESIS [J].
PAULUS, A ;
GASSMANN, E ;
FIELD, MJ .
ELECTROPHORESIS, 1990, 11 (09) :702-708
[22]   INSTABILITY OF THE MONOFUNCTIONAL ADDUCTS IN CIS-[PT(NH3)(2)(N7-N-METHYL-2-DIAZAPYRENIUM)CL](2+)-MODIFIED DNA - RATES OF CROSS-LINKING REACTIONS IN CIS-PLATINUM-MODIFIED DNA [J].
PAYET, D ;
GAUCHERON, F ;
SIP, M ;
LENG, M .
NUCLEIC ACIDS RESEARCH, 1993, 21 (25) :5846-5851
[23]   Reactions of the double-stranded oligonucleotide d(TTGGCCAA)(2) with cis-[Pt(NH3)(2)(H2O)(2)](2+) and [Pt(NH3)(3)(H2O)](2+) [J].
Reeder, F ;
Gonnet, F ;
Kozelka, J ;
Chottard, JC .
CHEMISTRY-A EUROPEAN JOURNAL, 1996, 2 (09) :1068-1076
[24]   Triammineplatinum(II) coordinated to a guanine does not prevent platination of an adjacent guanine in single-stranded oligonucleotides [J].
Reeder, F ;
Kozelka, J ;
Chottard, JC .
INORGANIC CHEMISTRY, 1996, 35 (05) :1413-&
[25]  
SATOW T, 1991, HRC-J HIGH RES CHROM, V14, P276
[26]   CAPILLARY ELECTROPHORETIC SEPARATION AND LASER-INDUCED FLUORESCENCE DETECTION OF THE MAJOR DNA-ADDUCTS OF CISPLATIN AND CARBOPLATIN [J].
SHARMA, M ;
JAIN, R ;
IONESCU, E ;
SLOCUM, HK .
ANALYTICAL BIOCHEMISTRY, 1995, 228 (02) :307-311
[28]   CISPLATIN AND DNA-REPAIR IN CANCER-CHEMOTHERAPY [J].
ZAMBLE, DB ;
LIPPARD, SJ .
TRENDS IN BIOCHEMICAL SCIENCES, 1995, 20 (10) :435-439
[29]  
ZEKANY L, 1992, ITERAT VERSION 2