Calcium independent receptor of alpha-latrotoxin,characterization and uses thereof
AssigneeNew York University
InventorsALEXANDRE G. PETRENKO, VALERY G. KRASNOPEROV
Life of this patent
Estimating the term…
Abstract
A novel receptor of α-latrotoxin (α-LTx) which binds α-LTx independently of calcium (Ca2+) presence and is thus a mediator of the calcium-independent stimulation of neurotransmitter release by α-latrotoxin has been isolated, purified and characterized. Designated CIRL (calcium-independent receptor of α-latrotoxin), it and its endogenous ligands can be used to modulate and regulate spontaneous calcium-independent neurotransmitter release and produce α-latrotoxin-like effects on the nerve terminal.
Claims44
A calcium-independent receptor of α-latrotoxin (CIRL) comprising a material selected from the group consisting of a protein, active fragments thereof, agonists thereof, mimics thereof, and combinations thereof, said receptor binding α-latrotoxin (α-LTx) both in the presence or absence of calcium.
An antibody to a calcium-independent receptor of α-latrotoxin (CIRL), the calcium-independent receptor of α-latrotoxin (CIRL) to which said antibody is raised binding α-latrotoxin (α-LTx) both in the presence or absence of calcium.
A DNA sequence or degenerate variant thereof, which encodes a calcium-independent receptor of α-latrotoxin (CIRL), or a fragment thereof, selected from the group consisting of:
- (A) the DNA sequence of FIG. 2B (SEQ ID NO:2);
- (B) DNA sequences that hybridize to any of the foregoing DNA sequences under standard hybridization conditions;
- and
- (C) DNA sequences that code on expression for an amino acid sequence encoded by any of the foregoing DNA sequences.
A recombinant DNA molecule comprising a DNA sequence or degenerate variant thereof, which encodes a calcium-independent receptor of α-latrotoxin (CIRL), or a fragment thereof, selected from the group consisting of:
- (A) the DNA sequence of FIG. 2B (SEQ ID NO:2);
- (B) DNA sequences that hybridize to any of the foregoing DNA sequences under standard hybridization conditions;
- and
- (C) DNA sequences that code on expression for an amino acid sequence encoded by any of the foregoing DNA sequences.
A host transformed with a recombinant DNA molecule comprising a DNA sequence or degenerate variant thereof, which encodes a calcium-independent receptor of α-latrotoxin (CIRL), or a fragment thereof, selected from the group consisting of:
- (A) the DNA sequence of FIG. 2B (SEQ ID NO:2);
- (B) DNA sequences that hybridize to any of the foregoing DNA sequences under standard hybridization conditions;
- and
- (C) DNA sequences that code on expression for an amino acid sequence encoded by any of the foregoing DNA sequences;
- wherein said DNA sequence is operatively linked to an expression control sequence.
A method for detecting the presence or activity of a calcium-independent receptor of α-latrotoxin (CIRL), said calcium-independent receptor of α-latrotoxin (CIRL) binding α-latrotoxin (α-LTx) both in the presence or absence of calcium;
- wherein said calcium-independent receptor of α-latrotoxin (CIRL) is measured by:
- A. contacting a biological sample from a mammal in which the presence or activity of said calcium-independent receptor of α-latrotoxin (CIRL) is suspected with a binding partner of said calcium-independent receptor of α-latrotoxin (CIRL) under conditions that allow binding of said calcium-independent receptor of α-latrotoxin (CIRL) to said binding partner to occur;
- and
- B. detecting whether binding has occurred between said calcium-independent receptor of α-latrotoxin (CIRL) from said sample and the binding partner;
- wherein the detection of binding indicates that presence or activity of said calcium-independent receptor of α-latrotoxin (CIRL) in said sample.
A method for detecting the presence and activity of a calcium-independent receptor of α-latrotoxin (CIRL) associated with a neurological disease in mammals comprising detecting the presence or activity of a calcium-independent α-latrotoxin receptor according to the method of claim 18, wherein detection of the presence or activity of the calcium-independent α-latrotoxin receptor indicates the presence and of a neurological or peripheral neuromuscular disease.
A method of testing the ability of a drug or other entity to modulate the activity of a calcium-independent receptor of α-latrotoxin (CIRL) which comprises
- A. culturing a colony of test cells which has the calcium-independent receptor of α-latrotoxin (CIRL) in a growth medium;
- B. adding the drug under test;
- and
- C. measuring the reactivity of said drug on said colony of test cells.
An assay system for screening drugs and other agents for ability to modulate the production and/or activity of a calcium-independent receptor of α-latrotoxin (CIRL), comprising:
- A. culturing an observable cellular test colony inoculated with a drug or agent;
- and
- B. examining said test colony for the presence of said calcium-independent receptor of α-latrotoxin (CIRL) wherein an increase or a decrease in the amount of said calcium-independent receptor of α-latrotoxin (CIRL) indicates the ability of a drug to modulate the production and/or the activity of said calcium-independent receptor of α-latrotoxin (CIRL), said calcium-independent receptor of α-latrotoxin (CIRL) binding α-latrotoxin (α-LTx) both in the presence or absence of calcium.
A test kit for the demonstration of a calcium-independent receptor of α-latrotoxin (CIRL) in a eukaryotic cellular sample, comprising:
- A. a predetermined amount of a detectably labelled specific binding partner of a calcium-independent receptor of α-latrotoxin (CIRL), said calcium-independent receptor of α-latrotoxin (CIRL) binding α-latrotoxin (α-LTx) both in the presence or absence of calcium;
- B. other reagents;
- and
- C. directions for use of said kit.
A method of preventing and/or treating cellular debilitations, derangements and/or dysfunctions and/or other disease states in mammals, comprising administering to a mammal a therapeutically effective amount of a material selected from the group consisting of a calcium-independent receptor of α-latrotoxin (CIRL), expressing cells thereof, an agent capable of promoting the production and/or activity of said calcium-independent receptor of α-latrotoxin (CIRL), an agent capable of mimicking the activity of said calcium-independent receptor of α-latrotoxin (CIRL), an agent capable of inhibiting the production of said calcium-independent receptor of α-latrotoxin (CIRL), and mixtures thereof, or a specific binding partner thereto, said calcium-independent receptor of α-latrotoxin (CIRL) binding α-latrotoxin (α-LTx) both in the presence or absence of calcium.
A pharmaceutical composition for the treatment of cellular debilitation, derangement and/or dysfunction in mammals, comprising:
- A. a therapeutically effective amount of a material selected from the group consisting of a calcium-independent receptor of α-latrotoxin (CIRL), expressing cells thereof, an agent capable of promoting the production and/or activity of said calcium-independent receptor of α-latrotoxin (CIRL), an agent capable of mimicking the activity of said calcium-independent receptor of α-latrotoxin (CIRL), an agent capable of inhibiting the production of said calcium-independent receptor of α-latrotoxin (CIRL), and mixtures thereof, or a specific binding partner thereto, said calcium-independent receptor of α-latrotoxin (CIRL) binding α-latrotoxin (α-LTx) both in the presence or absence of calcium;
- and
- B. a pharmaceutically acceptable carrier.
A calcium-independent receptor of α-latrotoxin (CIRL) implicated in neurological disturbances in mammals, said calcium-independent receptor of α-latrotoxin (CIRL) binding α-latrotoxin (α-LTx) both in the presence or absence of calcium.
A method of determining the calcium-independent receptor of α-latrotoxin (CIRL)-related pharmacological activity of a compound comprising:
- administering the compound to a mammal;
- determining the level and/or activity of calcium-independent receptor of α-latrotoxin (CIRL) proteins present;
- and
- comparing the level and/or activity of calcium-independent receptor of α-latrotoxin (CIRL) to a standard.
An antisense nucleic acid against a calcium-independent receptor of α-latrotoxin (CIRL) mRNA comprising a nucleic acid sequence hybridizing to said mRNA.
A recombinant DNA molecule having a DNA sequence which, on transcription, produces an antisense ribonucleic acid against a calcium-independent receptor of α-latrotoxin (CIRL) mRNA, said antisense ribonucleic acid comprising an nucleic acid sequence capable of hybridizing to said mRNA.
A ribozyme that cleaves calcium-independent receptor of α-latrotoxin (CIRL) mRNA.
Dependencies are inferred from the claim text.