US 2003/0198648 A1 · Methods for identifying compounds which inhibit binding of nucleocapsid 7 protein to HIV-1 RNA

Abstract

The present invention relates to methods of identifying a molecule from a library of molecules that inhibits binding of human immunodeficiency virus nucleocapsid 7 polypeptide (NCp7) to an oligonucleotide which comprises admixing an NCp7 polypeptide with at one labeled HIV-1 psi-site oligonucleotide and an amount of the molecule to be tested under binding conditions; and determining the amount of oligonucleotide bound to the NCp7 polypeptide, wherein a decrease in the amount of oligonucleotide bound in the presence of the molecule compared with the amount of oligonucleotide bound in the absence of the molecule indicates that the molecule inhibits binding of NCp7 polypeptide to the oligonucleotide.

Claims52

Dependent claims

A method for determining whether a compound inhibits formation of a complex between an HIV nucleocapsid protein 7 (NCp7) polypeptide and an HIV Ψ-site oligonucleotide comprising the steps of:

(a) admixing an NCp7 polypeptide with a compound;
(b) adding an HIV Ψ-site oligonucleotide to the admixture of step (a) under appropriate binding conditions so as to form an HIV Ψ-site oligonucleotide-NCp7 polypeptide complex;
and
(c) comparing the amount of complex formed in step (b) with the amount of complex formed in the absence of the compound, thereby determining whether the compound inhibits complex formation, wherein a decrease in the amount of complex formed in the presence of the compound indicates that the compound inhibits complex formation.

1 more element

A method for determining whether a compound inhibits binding of human immunodeficiency virus (HIV) nucleocapsid 7 polypeptide (NCp7) to an oligonucleotide which comprises:

(a) attaching an NCp7 polypeptide to a solid support, wherein the NCp7 polypeptide is selected from the group consisting of:
SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4;
(b) incubating the solid support with the NCp7 polypeptide linked thereto with a blocking agent;
(c) incubating the solid support with the NCp7 polypeptide linked thereto with:
(i) at least one labeled oligonucleotide selected from the group consisting of:
SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, a portion of a HIV-1 T-site RNA which retains an ability to bind to the NCp7 polypeptide, an oligonucleotide which binds to the NCp7 polypeptide under physiological conditions, and any combination thereof, and
(ii) an amount of the compound under binding conditions;
and
(d) determining the amount of labeled oligonucleotide bound to the NCp7 polypeptide, wherein a decrease in the amount of oligonucleotide bound in the presence of the compound compared with the amount of oligonucleotide bound in the absence of the compound indicates that the compound inhibits binding of NCp7 polypeptide to the oligonucleotide.

6 more elements

Dependencies are inferred from the claim text.

Methods for identifying compounds which inhibit binding of nucleocapsid 7 protein to HIV-1 RNA

Life of this patent

Methods for identifying compounds which inhibit binding of nucleocapsid 7 protein to HIV-1 RNA

Life of this patent

Abstract

Claims52