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http://dx.doi.org/10.5478/MSL.2015.6.1.7

Mass Spectrometric Determination of Zn2+ Binding/Dissociation Constant for Zinc Finger Peptides  

Lee, Choong Sik (Department of Chemistry, Sogang University)
Park, Soo Jin (Department of Chemistry, Sogang University)
Lee, Jae Young (Department of Life Science, Dongguk University)
Park, Sungsu (School of Mechanical Engineering, Sungkyunkwan University)
Jo, Kyubong (Department of Chemistry, Sogang University)
Oh, Han Bin (Department of Chemistry, Sogang University)
Publication Information
Mass Spectrometry Letters / v.6, no.1, 2015 , pp. 7-12 More about this Journal
Abstract
In the present study, we proposed a simple ESI-MS model for determining $Zn^{2+}$ binding (or dissociation) constants for zinc finger peptides (ZFPs) with a unique ${\beta}{\beta}{\alpha}$ fold consensus. The ionization efficiency (response) factors for this model, i.e., ${\alpha}$ and ${\beta}$, could be determined for ZiCo ZFP with a known $Zn^{2+}$ binding constant. We could determine the binding constants for other ZFPs assuming those with a ${\beta}{\beta}{\alpha}$ consensus conformation have the same ${\alpha}/{\beta}$ response ratio. In general, the ZPF dissociation constants exhibited $K_d$ values of $10^{-7}{\sim}10^{-9}M$, while $K_d$ values for a negative control non-specific $Zn^{2+}$ peptides were high, e.g., $5.5{\times}10^{-6}M$ and $4.3{\times}10^{-4}M$ for BBA1 and melittin, respectively.
Keywords
zinc finger; binding constant; electrospray-mass spectrometry; zinc ion;
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