Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-09-18
1999-06-22
Allen, Marianne P.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 691, 4353201, 435325, 435369, 536 235, 530350, C12N 1512, C07K 14705, G01N 3300
Patent
active
059142367
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
This invention relates to cloning and characterization of cellular receptors. Specifically, this invention relates to the cloning and characterization of the PCT-65 serotonin receptor protein.
BACKGROUND OF THE INVENTION
The neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) has a variety of functions in the central nervous system. It has been implicated in many cognitive and behavioral functions, including aggression, sexual behavior, learning and sleep. Disruptions of serotonergic systems may be a critical factor in a number of clinical disorders or conditions including schizophrenia, depression, obsessive compulsive disorder, anxiety, migraine headaches, and pain.
The multitude of effects produced by serotonin are mediated by various serotonin receptors which exist in the central and peripheral nervous system. The transduction of serotonergic signals across the neuronal membrane is believed to be mediated by a diversity of receptor subtypes which, in mammals, appear to fall into four pharmacologically distinct classes designated 5-HT.sub.1 -5-HT.sub.4. The 5HT.sub.1 subcategory has been further subdivided into five different subtypes referred to as 5HT.sub.1A-E. The primary structures for a number of these receptors have been elucidated by molecular cloning, including the 5-HT.sub.1, 5-HT.sub.2 and 5HT.sub.3 subclasses. In addition, the sequences of three different Drosophila serotonin receptors, 5-HT.sub.dro1 and 5-HT.sub.dro2A,B, have been reported.
Selective therapeutic agents, including agonist and antagonist drugs, have been developed based on serotonin receptor technologies utilizing the serotonin classes known in the art. 5-HT.sub.2 antagonists, for example, are useful in the treatment of schizophrenia, parkinsonism, and anxiety disorders. Several azapirones, such as buspirone, gepirone, and ipsapirone, have high affinities for 5HT.sub.1A receptors in the brain, and are useful in the treatment of anxiety. Highly selective 5-HT uptake inhibitors, which have minimal effects on norepinephrine or dopamine uptake or on other neurotransmitter receptors, have been used to successfully treat depression.
Characterization of proteins with serotonin receptor activity would clarify the role of serotonin in the central nervous system. Analysis of the receptor proteins and their functional role in the central nervous system would help elucidate the pathophysiological basis of many human diseases. Accordingly, disclosed herein is a structurally and pharmacologically novel serotonin receptor which is distinct from any class of previously described 5-HT receptors.
BRIEF DESCRIPTION OF THE FIGURES
FIGS. 1A-B are photographs of Northern blots of rat Poly (A).sup.+ RNA identifying PCT-65 mRNA from various central nervous system (CNS) and peripheral tissues. RNA size markers (in kilobases) is provided at the right of each blot.
FIG. 2 illustrates the cloning procedure used to derive the PCT-65 clone without an intron for expression studies.
FIG. 3a-1 provides the results of binding studies to assess the ability of PCT-65 containing construct. FIG. 3a-2 additionally includes a Scatchard analysis of the saturation data. FIG. 3b illustrates the pharmacological presence of various serotonin agonists and antagonists as well as the effect of PCT-65 receptor modulation using Gpp(NH)p.
SUMMARY OF THE INVENTION
One embodiment of the present invention is the isolated mammalian serotonin receptor protein PCT-65. Preferably this receptor protein is human. The present invention also encompasses species variations of the PCT-65 receptor.
Another embodiment of the present invention is a method for screening a drug candidate for central nervous system activity by contacting the drug candidate with the PCT-65 protein and measuring binding of the drug candidate by the protein.
An additional embodiment of the present invention is a method for screening drug candidate for central nervous system activity by first contacting the PCT-65 serotonin receptor protein with a first molecule known to be bound
REFERENCES:
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Shen, Y., et al., (1993) Molecular cloning and expression of a 5-Hydroxytryptamine.sub.7 . . . Journal of Biological Chemistry 268(24):18200-18204.
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Hamblin Mark
Monsma, Jr. Frederick J.
Shen Yong
Sibley David R.
Allen Marianne P.
United States of America
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