Prev. Article Next Article Table of Contents

Article

Selective Recognition of a DNA G-Quadruplex by an Engineered Antibody^†

Supporting Info

Himesh Fernando, Rapha

l Rodriguez and Shankar Balasubramanian*

The University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, U.K.

Biochemistry, 2008, 47 (36), pp 9365–9371

DOI: 10.1021/bi800983u

Publication Date (Web): August 15, 2008

†

This research was supported by a programme grant from Cancer Research UK. H.F. was funded by Cambridge Commonwealth Trusts.

* To whom correspondence should be addressed. Tel: (+) 44 1223 336 347. Fax: (+) 44 1223 336 913. E-mail: sb10031@cam.ac.uk.

Abstract

Particular guanine rich nucleic acid sequences can fold into stable secondary structures called G-quadruplexes. These structures have been identified in various regions of the genome that include the telomeres, gene promoters and UTR regions, raising the possibility that they may be associated with biological function(s). Computational analysis has predicted that intramolecular G-quadruplex forming sequences are prevalent in the human genome, thus raising the desire to differentially recognize genomic G-quadruplexes. We have employed antibody phage display and competitive selection techniques to generate a single-chain antibody that shows >1000-fold discrimination between G-quadruplex and duplex DNA, and furthermore >100-fold discrimination between two related intramolecular parallel DNA G-quadruplexes. The amino acid sequence composition at the antigen binding site shows conservation within the light and heavy chains of the selected scFvs, suggesting sequence requirements for G-quadruplex recognition. Circular dichroism (CD) spectroscopic data showed that the scFv binds to the prefolded G-quadruplex and does not induce G-quadruplex structure formation. This study demonstrates the strongest discrimination that we are aware of between two intramolecular genomic G-quadruplexes.

View: Full Text HTML | Hi-Res PDF | PDF w/ Links

Tools

SciFinder Links

Get Reference Detail

History

Published In Issue September 09, 2008
Article ASAPAugust 15, 2008
Received: May 23, 2008
Revised: July 10, 2008

Recommend & Share

Related Content

A Novel Small Molecule That Alters Shelterin Integrity and Triggers a DNA-Damage Response at TelomeresJournal of the American Chemical Society
- A Novel Small Molecule That Alters Shelterin Integrity and Triggers a DNA-Damage Response at Telomeres
  Raphal Rodriguez, Sebastian Mller, Justin A. Yeoman, Chantal Trentesaux, Jean-Franois Riou and Shankar Balasubramanian
  Journal of the American Chemical Society 2008 130 (47), pp 15758–15759
  Abstract: We describe a novel synthetic small molecule which shows an unprecedented stabilization of the human telomeric G-quadruplex with high selectivity relative to double-stranded DNA. We report that this compound can be used in vitro to inhibit telomerase ...
  Abstract | Full Text HTML | PDF w/ Links | Hi-Res PDF
Diarylethynyl Amides That Recognize the Parallel Conformation of Genomic Promoter DNA G-QuadruplexesJournal of the American Chemical Society
- Diarylethynyl Amides That Recognize the Parallel Conformation of Genomic Promoter DNA G-Quadruplexes
  Jyotirmayee Dash, Pravin S. Shirude, Shang-Te Danny Hsu and Shankar Balasubramanian
  Journal of the American Chemical Society 2008 130 (47), pp 15950–15956
  Abstract: We report bis-phenylethynyl amide derivatives as a potent G-quadruplex binding small molecule scaffold. The amide derivatives were efficiently prepared in 3 steps by employing Sonogashira coupling, ester hydrolysis and a chemoselective amide coupling. ...
  Abstract | Full Text HTML | PDF w/ Links | Hi-Res PDF
Rational Design of Substituted Diarylureas: A Scaffold for Binding to G-Quadruplex MotifsJournal of Medicinal Chemistry
- Rational Design of Substituted Diarylureas: A Scaffold for Binding to G-Quadruplex Motifs
  William C. Drewe, Rupesh Nanjunda, Mekala Gunaratnam, Monica Beltran, Gary N. Parkinson, Anthony P. Reszka, W. David Wilson and Stephen Neidle
  Journal of Medicinal Chemistry 2008 51 (24), pp 7751–7767
  Abstract: The design and synthesis of a series of urea-based nonpolycyclic aromatic ligands with alkylaminoanilino side chains as telomeric and genomic G-quadruplex DNA interacting agents are described. Their interactions with quadruplexes have been examined by ...
  Abstract | Full Text HTML | PDF w/ Links | Hi-Res PDF

Article

Selective Recognition of a DNA G-Quadruplex by an Engineered Antibody^†