Biophysical characterization of VEGF-aHt DNA aptamer interactions

Indhu Kanakaraj; Wen-Hsiang Chen; Mohan Poongavanam; Sagar Dhamane; Loren J Stagg; John E Ladbury; Katerina Kourentzi; Ulrich Strych; Richard C Willson

doi:10.1016/j.ijbiomac.2013.02.016

Biophysical characterization of VEGF-aHt DNA aptamer interactions

Int J Biol Macromol. 2013 Jun:57:69-75. doi: 10.1016/j.ijbiomac.2013.02.016. Epub 2013 Mar 5.

Authors

Indhu Kanakaraj¹, Wen-Hsiang Chen, Mohan Poongavanam, Sagar Dhamane, Loren J Stagg, John E Ladbury, Katerina Kourentzi, Ulrich Strych, Richard C Willson

Affiliation

¹ Department of Biology and Biochemistry, University of Houston, TX, USA.

PMID: 23470436
DOI: 10.1016/j.ijbiomac.2013.02.016

Abstract

The binding of the well-studied DNA aptamer aHt (5'-ATACCAGTCTATTCAATTGGGCCCGTCCGTAT GGTGGGTGTGCTGGCCAG-3'), which has been demonstrated to recognize human vascular endothelial growth factor (VEGF₁₆₅) to recombinant VEGF was characterized using fluorescence anisotropy, isothermal titration calorimetry and analytical ultracentrifugation. The negatively-charged DNA aptamer is selective for VEGF and does not recognize positively-charged hen egg lysozyme, or bovine serum albumin. In contrast to the VEGF association of the previously-described aV DNA aptamer, where the binding is enthalpically driven and sequence-specific, the binding of the aHt aptamer to VEGF is entropically-driven and not abolished by scrambling of the sequence.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Aptamers, Nucleotide / chemical synthesis
Aptamers, Nucleotide / chemistry*
Cattle
Chickens
Humans
Muramidase / chemistry
Serum Albumin, Bovine / chemistry
Vascular Endothelial Growth Factor A / antagonists & inhibitors
Vascular Endothelial Growth Factor A / chemistry*

Substances

Aptamers, Nucleotide
VEGFA protein, human
Vascular Endothelial Growth Factor A
Serum Albumin, Bovine
Muramidase