Methods for detecting and quantifying binding and inhibition of binding of species to nucleic acids
AssigneeMessage Pharmaceuticals
InventorsJack Summers, Michael Sturgess, John Shimko
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Abstract
The invention features methods for detecting and quantifying the binding or inhibition of binding of species to biopolymers, e.g., nucleic acids. The invention is based on the use of probes that have magnetic relaxation properties that are affected by the presence of paramagnetic metal ions, e.g., Mn2+. Any class of biopolymer that binds metal ions at its active site or uses metal ion cofactors can be studied using these methods.
Claims41
A method of detecting the binding of a species to a nucleic acid, said method comprising the steps of:
- (a) measuring a magnetic relaxation property of a probe in a first solution using nuclear magnetic resonance spectroscopy, wherein said first solution comprises said nucleic acid, a first concentration of said species, a paramagnetic metal ion, and said probe;
- (b) measuring the magnetic relaxation property of said probe in a second solution using nuclear magnetic resonance spectroscopy, wherein said second solution comprises said nucleic acid, a second concentration of said species, a paramagnetic metal ion, and said probe;
- and
- (c) comparing the relaxation property measured in step (a) with the relaxation property measured in step (b), wherein a difference in said relaxation properties indicates the binding of said species to said nucleic acid.
A method of detecting the binding of a paramagnetic metal ion to a nucleic acid, said method comprising the steps of:
- (a) measuring a magnetic relaxation property of a probe in a first solution using nuclear magnetic resonance spectroscopy, wherein said first solution comprises a first concentration of said paramagnetic metal ion, said nucleic acid, and said probe;
- (b) measuring the magnetic relaxation property of said probe in a second solution using nuclear magnetic resonance spectroscopy, wherein said second solution comprises a second concentration of said paramagnetic metal ion, said nucleic acid, and said probe;
- and
- (c) comparing the relaxation property measured in step (a) with the relaxation property measured in step (b), wherein a difference in said relaxation properties indicates the binding of said paramagnetic metal ion to said nucleic acid.
A method of determining the effect of a first species on the binding of a second species to a nucleic acid, said method comprising the steps of:
- (a) measuring a magnetic relaxation property of a probe in a first solution using nuclear magnetic resonance spectroscopy, wherein said first solution comprises a first concentration of said first species, said nucleic acid, said second species that binds to said nucleic acid, a paramagnetic metal ion, and a probe;
- (b) measuring the magnetic relaxation property of said probe in a second solution using nuclear magnetic resonance spectroscopy, wherein said second solution comprises a second concentration of said first species, said nucleic acid, said second species, said paramagnetic metal ion, and said probe;
- and
- (c) comparing the relaxation property measured in step (a) with the relaxation property measured in step (b), wherein a difference in said relaxation properties indicates the inhibition of the binding of said second species to said nucleic acid.
The method of claim 31, wherein said metal ion is Mg2+.
A method of detecting the availability of a paramagnetic metal ion in a solution comprising a nucleic acid, said method comprising the steps of:
- (a) providing a solution comprising said nucleic acid, a probe, and said paramagnetic metal ion and
- (b) measuring a magnetic relaxation property of said probe in said solution using nuclear magnetic resonance spectroscopy, wherein the magnitude of said magnetic property is indicative of the availability of said paramagnetic ion in said solution.
Dependencies are inferred from the claim text.