α-Hemolysin nanopore studies reveal strong interactions between biogenic polyamines and DNA hairpins

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• 作者：Yun Ding ; Aaron M. Fleming ; Cynthia J. Burrows
• 关键词：α ; Hemolysin ; DNA hairpin ; Spermine ; Spermidine ; Putrescine
• 刊名：Microchimica Acta
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：183
• 期：3
• 页码：973-979
• 全文大小：1,711 KB
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• 作者单位：Yun Ding (1)
  Aaron M. Fleming (1)
  Cynthia J. Burrows (1)
  
  1. Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, UT, 84112-0850, USA
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Inorganic Chemistry
  Physical Chemistry
  Characterization and Evaluation Materials
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Wien
• ISSN：1436-5073

文摘

The α-hemolysin (α-HL) nanopore is capable of analyzing DNA as it is electrophoretically driven through the pore. Respective current vs. time (i-t) traces depend on the DNA sequence, its secondary structures, or on the physical conditions of the analysis. The current study describes the analysis of a DNA hairpin with a 5′-extension by applying α-HL nanopores in the presence of the polyamines spermine (Spm), spermidine (Spd), and putrescine (Put) and revealed i-t traces characteristic of the DNA-polyamine complex. Voltage-dependent studies also revealed that the hairpin-Spm complex formed with excess Spm cannot be unzipped and translocated through the pores even if the voltage is increased to 180 mV. The DNA hairpin sample was titrated with Spm, Spd, or Put that showed a dose-dependent response in the characteristic event patterns for hairpins bound to Spm or Spd, but not for Put. Plots of the event types vs. counts were used to calculate binding constants for the Spm or Spd hairpin interactions. The titration also demonstrated that the event rate decreased ~10-fold on increasing the Spm or Spd concentrations from 0 to 4 mM. These observations impose practical limitations on the ability to use Spm or Spd for DNA studies with the α-HL nanopore.