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Characterization of noncovalent complexes formed between minor groove binding molecules and duplex DNA by electrospray ionization mass spectrometry
被引:98
作者:
Gale, DC
[1
]
Smith, RD
[1
]
机构:
[1] PACIFIC NW LAB, DEPT CHEM SCI, CHEM METHODS & SEPARAT GRP, RICHLAND, WA 99352 USA
关键词:
D O I:
10.1016/1044-0305(95)00530-7
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
The noncovalent complex formed in solution between minor groove binding molecules and an oligonucleotide duplex was investigated by electrospray ionization-mass spectrometry (ESI-MS). The oligonucleotide duplex formed between two sequence-specific 14-base pair oligonucleotides was observed intact by ESI-MS and in relatively high abundance compared to the individual single-stranded components. Only sequence-specific A:B duplexes were observed, with no evidence of random nonspecific aggregation (i.e., A:A or B:B) occurring under the conditions utilized. Due to the different molecular weights of the two 14-base Fair oligonucleotides, unambiguous determination of each oligonucleotide and the sequence-specific duplex was confirmed through their detection at unique mass-to-charge ratios. The noncovalent complexes formed between the self-complementary 5'-dCGCAAA7TTTGCG-3' oligonucleotide and three minor groove binding molecules (distamycin A, pentamidine, and Hoechst 33258) were also observed. Variation of several electrospray ionization interface parameters as well as collision-induced dissociation methods were utilized to characterize the nature and stability of the noncovalent complexes. The noncovalent complexes upon collisional activation dissociated into single-stranded oligonucleotides and single-stranded oligonucleotides associated with a minor groove binding molecule. ESI-MS shows potential for the study of small molecule-oligonucleotide duplex interactions and determination of small molecule binding stoichiometry.
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页码:1154 / 1164
页数:11
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