Towards zirconium phosphonate-based microarrays for probing DNA-protein interactions: critical influence of the location of the probe anchoring groups.
Terminal phosphate groups on double-stranded DNA probes bind strongly to glass substrates coated with a zirconium phosphonate monolayer, and probes immobilized in this way as microarrays can be used to detect protein targets. The sensitivity of the microarray was shown to be enhanced by the use of a polyguanine segment ((G)n , n > or = 5) as a spacer between the phosphate linker and the protein interaction domain. More importantly, the presence of phosphate linkers on both ends of the dsDNA probes leads to significant enhancement of target capture. The relevant characteristics of the different probes when bound to the surface were determined, by the original use of a combination of surface characterization techniques (XPS, AFM, and Sarfus). In this context, the location of the phosphate linkers in the duplex probes was found to result in different probe surface coverage and presentation on the surface, which affect subsequent interactions with the target protein.
Monot
Julien
J
Université de Nantes, CNRS, UMR 6230, Chimie Et Interdisciplinarité: Synthèse Analyse Modélisation, UFR Sciences et Techniques, 2, rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
Petit
Marc
M
Lane
Sarah M
SM
Guisle
Isabelle
I
Léger
Jean
J
Tellier
Charles
C
Talham
Daniel R
DR
Bujoli
Bruno
B
D000327:Adsorption; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D002942:Circular Dichroism; D003432:Cross-Linking Reagents / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry*; D063065:Organophosphonates / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics; D013499:Surface Properties; D046949:Thermotoga neapolitana / Q000235:genetics / Q000378:metabolism; D015040:Zirconium / Q000737:chemistry*
C087018:ArgR protein, Bacteria; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D015342:DNA Probes; D015345:Oligonucleotide Probes; D063065:Organophosphonates; D012097:Repressor Proteins; D015040:Zirconium