Parallel, Quantitative Measurement of Protein Binding to a 120-Element Double-Stranded DNA Array in Real Time Using Surface Plasmon Resonance Microscopy

Parallel, Quantitative Measurement of Protein Binding to a 120-Element Double-Stranded DNA Array in Real Time Using Surface Plasmon Resonance Microscopy

Quantitative, real-time measurement of kinetics of sequence-specific binding of DNA-binding proteins to double-stranded DNA (dsDNA) immobilized in a 10 × 12 array on a planar gold surface is demonstrated using surface plasmon resonance (SPR) microscopy. This binding of the yeast transcription factor...

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Veröffentlicht in:Analytical chemistry (Washington) 2004-04, Vol.76 (7), p.2071-2082
Hauptverfasser: Shumaker-Parry, Jennifer S, Aebersold, Ruedi, Campbell, Charles T
Format: Artikel
Sprache:eng
Schlagworte:
Adsorption
Analytical biochemistry: general aspects, technics, instrumentation
Analytical, structural and metabolic biochemistry
Animals
Base Sequence
Binding sites
Biological and medical sciences
Deoxyribonucleic acid
Desorption
DNA
DNA - chemistry
DNA - metabolism
DNA-Binding Proteins - analysis
Fundamental and applied biological sciences. Psychology
Gold - chemistry
Gold - pharmacology
Hydrogen-Ion Concentration
Kinetics
Microscopy
Molecular Sequence Data
Oligonucleotide Array Sequence Analysis - methods
Proteins
Surface Plasmon Resonance - methods
Surface Properties
Time Factors
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Zusammenfassung:Quantitative, real-time measurement of kinetics of sequence-specific binding of DNA-binding proteins to double-stranded DNA (dsDNA) immobilized in a 10 × 12 array on a planar gold surface is demonstrated using surface plasmon resonance (SPR) microscopy. This binding of the yeast transcription factor Gal4 to a 120-spot dsDNA array made with alternating 200-μm spots of its dsDNA operator sequence and an unrelated DNA sequence proves that this method could be used to simultaneously monitor the kinetics of binding of proteins to 120 different dsDNA sequences with a sensitivity to ∼0.5 pg (
ISSN:0003-2700
1520-6882
DOI:10.1021/ac035159j