Iron-responsive element-binding protein (IRP) is a cytosolic protein which binds to iron-responsive element (IRE)-mRNA.1,2 Recent report indicated that Mn2+ ion can mimic Fe+2, which plays a significant role for IRPs by interfering with the binding between IRE-RNAs and the IRPs.3 Because surface plasmon resonance (SPR) provides an optical method to interrogate systems with submonolayer sensitivity, an SPR study of the IRE-Mn2+ ion system has been performed. We have prepared a 77nt RNA which contains the IRP binding loop1 and a spacer for surface immobilization. The IRE construct is hybridized to a short complementary DNA sequence end modified with a multi-thiol dendrimer4 in order to immobilize the IRE-RNA on the SPR detection gold surface. This multi-thiol modification of ssDNA molecules is much more robust5 and have wide range applications such as directed assembly of proteins.6 Our preliminary SPR results indicate that the binding of Mn2+ ion to IRE-RNA is observable via SPR. An extension of this study will be the in vitro determination of the kinetics of IRP association and dissociation with IRE's using SPR.  Reference:  (1)   Walden, W. E.; Selezneva, A. I.; Dupuy, J.; Volbeda, A.; Fontecilla-Camps, J. C.; Theil, E. C.; Volz, K. Science 2006 , 314, 1903.(2)   Ma, J.; Haldar, S.; Khan, M. A.; Sharma, S. D.; Merrick, W. C.; Theil, E. C.; Goss, D. J. Proc Natl Acad Sci U S A 2012 , 109, 8417.(3)   Khan, M. A.; Walden, W. E.; Goss, D. J.; Theil, E. C. J Biol Chem 2009 , 284, 30122.(4)   Day, B. S.; Fiegland, L. R.; Vint, E. S.; Shen, W.; Morris, J. R.; Norton, M. L. Langmuir 2011 , 27, 12434.(5)   Herne, T. M.; Tarlov, M. J. Journal of the American Chemical Society 1997 , 119, 8916.(6)   Boozer, C.; Ladd, J.; Chen, S.; Jiang, S. Analytical Chemistry 2006 , 78, 1515.

Surface plasmon resonance detection of Mn<sup>2+</sup> binding to iron responsive element-mRNA

246th National Meeting (2013)

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Surface plasmon resonance detection of Mn2+ binding to iron responsive element-mRNA

Surface plasmon resonance detection of Mn²⁺ binding to iron responsive element-mRNA