Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
2000-06-01
2003-07-01
Spector, Lorraine (Department: 1647)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S252300, C435S320100, C536S023500
Reexamination Certificate
active
06586205
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to novel GPCR-like nucleic acid sequences and proteins. Also provided are vectors, host cells, and recombinant methods for making and using the novel molecules.
BACKGROUND OF THE INVENTION
G-protein coupled receptors (GPCRs) constitute a major class of proteins responsible for transducing a signal within a cell. GPCRs have three structural domains: an amino terminal extracellular domain, a transmembrane domain containing seven transmembrane segments, three extracellular loops, and three intracellular loops, and a carboxyl terminal intracellular domain. Upon binding of a ligand to an extracellular portion of a GPCR, a signal is transduced within the cell that results in a change in a biological or physiological property of the cell. GPCRs, along with G-proteins and effectors (intracellular enzymes and channels modulated by G-proteins), are the components of a modular signaling system that connects the state of intracellular second messengers to extracellular inputs.
GPCR genes and gene-products are potential causative agents of disease (Spiegel et al (1993),
J. Clin. Invest
. 92:1119-1125; McKusick et al.,
J. Med. Genet
. (1993) 30:1-26). For example, specific defects in the rhodopsin gene and the V2 vasopressin receptor gene have been shown to cause various forms of retinitis pigmentosum (Nathans et al. (1992)
Annu. Rev. Genet
. 26:403-424), and nephrogenic diabetes insipidus (Holtzman et al. (1993)
Hum. Mol. Genet
. 2:1201-1204). These receptors are of critical importance to both the central nervous system and peripheral physiological processes. Evolutionary analyses suggest that the ancestor of these proteins originally developed in concert with complex body plans and nervous systems.
Metabotrophic (P2Y) receptors form a distinct subset of G-protein coupled receptors, whose ligands comprise adenosine 5′-triphosphate (ATP) and/or related nucleotides. P2Y receptors are generally distinguished pharmacologically by the rank order of effectiveness of agonists: some prefer pyrimidines to purines. More than eleven P2Y receptors have been reported. For a review see, for example, North et al. (1997)
Current Opinion in Neurobiology
7:346-357. Presently, four subfamilies have been distinguished: 1) P2Y
1
, 2) P2Y
2
/P2Y
4
/P2Y
8
, 3) P2Y
3
/P2Y
6
, and, 4) P2Y
5
. The P2 receptors may be coupled through G-proteins to signaling pathways involving phospholipase C (which increases inositol-1,4,5-trisphosphate and diacylglycerol formation), phospholipase A2 (with consequent generation of eicosanoids), or adenylate cyclase (which increases cAMP levels). Some members of this receptor family have also been shown to mediate their signals through the inhibition of adenylate cyclase, the inhibition of N-type Ca
2+
channels, and the activation of K
+
channels. For a review see Boarder et al. (1995)
Trends Pharmacol Sci
16:133-139 and Neary et al. 1996
Trends Neurosci
19:13-18.
Endogenous ATP as well as the uracil congener, UTP, act as extracellular signaling molecules and mediate some of their effects via interactions through various members of the P2Y receptor family. Signal transduction through P2Y receptors has recently been implicated in the modulation of neuronal cell membrane ion channels. P2Y type receptors in the rat cerebella or hippocampal neurons (Ikeuchi et al. (1996)
Biochem Biophys Res Commun
218:67-71) and guinea pig atrial cells (Matsuura et al. (1996)
J Physiol
490:659-671) have been linked to the activation of K
+
channels. This coupling is G-protein mediated and membrane-delimited, thus adding P2Y receptors to the family of seven-transmembrane receptors known to act in this way. Furthermore, nucleotides inhibit endogenous Ca
2+
currents in neuroblastoma hybrid cells, which contain the P2Y
2
receptor. The direct involvement of P2Y
2
receptors in this cellular event has since been demonstrated through a direct approach. Rat sympathetic neurons have no native response to UTP. Microinjection of P2Y
2
receptor cRNA into the rat sympathetic neurons resulted in expression of the P2Y
2
receptor and an acquired UTP-mediated inhibition of their N-type Ca
2+
channels (Chen et al. (1996)
Endocrinology
137:1833-1840 and Nicholas et al. (1996)
Mol Pharmacol
50:224-229). Since P2Y
2
receptors are also known to activate phospholipase C, these results indicate that P2Y
2
receptors can mediate signal transduction through two independent pathways, depending on the cellular environment.
In P2Y
1
-transfected COS-7 cells, agonists produce a transient increase in internal Ca
2+
associated with the formation of inositol-1,4,5-trisphosphate (Simon et al. (1995)
Eur J Pharmacol
291:281-289). P2Y
1
receptors have been cloned from turkey brain, mouse and rat insulinoma cells, bovine aortic endothelia cells, rat brain, and human erythroid leukemia cells. Interestingly, the rat P2Y
1
receptors are expressed in endothelial cells (B10) isolated from the blood brain barrier. Various nucleotides have been found to mobilize Ca
2+
in these cells (Webb et al. (1996)
J Pharmacol
119:1385-1392). However, the Ca
2+
mobilization response is associated not with an increase in inositol-1,4,5-trisphosphate but rather with the inhibition of adenylate cyclase. This is in contract to the signal transduction mediated by turkey (Filtz et al. (1994)
Mol Pharmacol
46:8-14), chicken (Simon et al. (1995)
Pharmacol Toxicol
76:302-307) or human (Schachter et al. (1 996)
Br J Phrmacol
118:167-173) P2Y
1
receptors which when expressed heterologously, clearly induced only inositol-1,4,5-trisphosphate formation. Hence, the P2Y
1
receptor, while commonly coupled to phospholipase C, can in some native cells be coupled instead through the G
0
/G
i
cyclase inhibitory pathway.
As extracellular signaling molecules, ATP and UTP are involved in various physiological and pathophysiological processes that have been associated with P2Y receptor family members. The role of ATP in tissue homeostasis, fast excitatory neurotransmission, tissue development, pain transmission, macrophage apoptosis, platelet aggregation, astroglia cell function, and the development and maturation of the nervous system has been established and current evidence indicates that the P2Y receptors mediate many of these cellular effects via an interaction with the extracellular ATP ligand. See for example, Burnstock et al. (1996)
Drug Dev. Res
. 39:204-242 and Williams et al (1999)
Progress in Brain Research
120:93-106. Furthermore, UTP, acting via the P2Y receptors, is a potent and selective modulator of mucocilliary transport. Thus, members of the P2Y receptor family mediate cellular responses produced by extracellular ATP, UTP, and/or related nucleotides (Anderson et al. (1997)
Trends Pharmacol. Sci
. 18:387-392).
Extracellular ATP is known to be hyperalgesic: peripheral administration of ATP produces pain and ATP can enhance the production of prostaglandins that also produce pain (Needleman et al. (1974)
Circ. Res
. 34:455-460). Burnstock ((1996)
Autonomic Neuroscience Institute
, sheet 8) has suggested that locally produced ATP may contribute to the pain associated with causalgia, reflex sympathetic dystrophy, migraine, angina, lumbar, pelvic and cancer pain. Salter et al. have shown that P2Y receptors in dorsal horn astrocytes respond to ATP by increasing Ca
2+
levels and thus, P2Y receptors may be involved in mediating the pain response (Salter et al. (1994)
J. Neurosci
. 14:1563-1575). During reactive-hyperemia, large amounts of ATP are released from vascular endothelial cells that act on P2Y receptors, resulting in the release of nitric oxide and vasodilatation. Burnstock proposed that in the microcirculation, ATP diffuses from the endothelial cells to activate nociceptive endings of sensory nerve fibers in the adventitia (Burnstock (1996)
Lancet
347:1604-1605). ATP released from platelets during aggregation, which has been reported to increase in migraine, may also contribute to the initiation of pain via nocicepti
Glucksmann Maria Alexandra
Silos-Santiago Inmaculada
Millennium Pharmaceuticals Inc.
Millennium Pharmaceuticals Inc.
Spector Lorraine
Wegert Sandra
LandOfFree
43239 a novel GPCR-like molecule and uses thereof does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with 43239 a novel GPCR-like molecule and uses thereof, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and 43239 a novel GPCR-like molecule and uses thereof will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3046309