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

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

Houston Methodist

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

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.

Original language	English (US)
Pages (from-to)	69-75
Number of pages	7
Journal	International Journal of Biological Macromolecules
Volume	57
DOIs	https://doi.org/10.1016/j.ijbiomac.2013.02.016
State	Published - Jun 2013

Keywords

Aptamer
Fluorescence anisotropy
Isothermal titration calorimetry
Vascular endothelial growth factor

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Structural Biology

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10.1016/j.ijbiomac.2013.02.016

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title = "Biophysical characterization of VEGF-aHt DNA aptamer interactions",

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.",

keywords = "Aptamer, Fluorescence anisotropy, Isothermal titration calorimetry, Vascular endothelial growth factor",

author = "Indhu Kanakaraj and Chen, {Wen Hsiang} and Mohan Poongavanam and Sagar Dhamane and Stagg, {Loren J.} and Ladbury, {John E.} and Katerina Kourentzi and Ulrich Strych and Willson, {Richard C.}",

note = "Funding Information: This research was funded in part by grants to RCW from the Welch foundation ( Grant E-1264 ), NASA and NSF .",

year = "2013",

month = jun,

doi = "10.1016/j.ijbiomac.2013.02.016",

language = "English (US)",

volume = "57",

pages = "69--75",

journal = "International Journal of Biological Macromolecules",

issn = "0141-8130",

publisher = "Elsevier B.V.",

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T1 - Biophysical characterization of VEGF-aHt DNA aptamer interactions

AU - Kanakaraj, Indhu

AU - Chen, Wen Hsiang

AU - Poongavanam, Mohan

AU - Dhamane, Sagar

AU - Stagg, Loren J.

AU - Ladbury, John E.

AU - Kourentzi, Katerina

AU - Strych, Ulrich

AU - Willson, Richard C.

N1 - Funding Information: This research was funded in part by grants to RCW from the Welch foundation ( Grant E-1264 ), NASA and NSF .

PY - 2013/6

Y1 - 2013/6

N2 - 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.

AB - 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.

KW - Aptamer

KW - Fluorescence anisotropy

KW - Isothermal titration calorimetry

KW - Vascular endothelial growth factor

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DO - 10.1016/j.ijbiomac.2013.02.016

M3 - Article

C2 - 23470436

AN - SCOPUS:84875790899

SN - 0141-8130

VL - 57

SP - 69

EP - 75

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

ER -

Biophysical characterization of VEGF-aHt DNA aptamer interactions

Abstract

Keywords

ASJC Scopus subject areas

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