Life Sciences 78 (2005) 506 – 511 www.elsevier.

com/locate/lifescie

Minireview

Screening the receptorome for plant-based psychoactive compounds
Kerry Ann O’Connor a, Bryan L. Roth a,b,c,*
a b

Department of Biochemistry, Case Western Reserve University, Cleveland, OH, USA Department of Neurosciences, Case Western Reserve University, Cleveland, OH, USA c Department of Psychiatry, Case Western Reserve University, Cleveland, OH, USA

Abstract Throughout time, humans have used psychoactive plants and plant-derived products for spiritual, therapeutic and recreational purposes. Furthermore, the investigation of psychoactive plants such as Cannabis sativa (marijuana), Nicotiana tabacum (tobacco) and analogues of psychoactive plant derivatives such as lysergic acid diethylamide (LSD) have provided insight into our understanding of neurochemical processes and diseases of the CNS. Currently, many of these compounds are being used to treat a variety of diseases, such as depression and anxiety in the case of Piper methysticum Kava Kava (Martin et al., 2002; Singh and Singh, 2002). G-protein coupled receptors (GPCRs) are the most common molecular target for both psychoactive drugs and pharmaceuticals. The ‘‘receptorome’’ (that portion of the genome encoding ligand reception) encompasses more than 8% of the human genome (Roth et al., 2004) and as such provides a large number of possible targets for psychoactive drug interactions. A systematic, comprehensive study is necessary to identify novel active psychoactive plant-based compounds and the molecular targets of known compounds. Herein we describe the development of a high throughput system (HTS) to screen psychoactive compounds against the receptorome and present two examples (Salvia divinorum, the ‘‘magic mint’’ hallucinogen and Banisteriopsis caapi, the main component of Ayahuasca, a psychoactive beverage) where HTS enabled the identification of the molecular target of each compound. D 2005 Elsevier Inc. All rights reserved.
Keywords: GPCR; Salvinorin A; Ayahuasca

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . The receptorome . . . . . . . . . . . . . . . . . . . . . . . . . Receptoromics . . . . . . . . . . . . . . . . . . . . . . . . . . Virtual screening of the receptorome . . . . . . . . . . . . . Physical screening of the receptorome. . . . . . . . . . . . . Molecular targets of naturally occurring psychoactive-compounds Salvia divinorum . . . . . . . . . . . . . . . . . . . . . . . Banisteriopsis caapi . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 506 507 507 507 508 508 508 509 509 509

Introduction Psychoactive plants exert profound effects on human consciousness, emotion, and cognition, and as such have been
* Corresponding author. Department of Biochemistry, Room W441, Case Western Reserve University School of Medicine, 2109 Adelbert Road, Cleveland, OH 44106-4935, USA. Tel.: +1 216 368 2730; fax: +1 216 368 3419. E-mail address: bryan.roth@case.edu (B.L. Roth). 0024-3205/$ - see front matter D 2005 Elsevier Inc. All rights reserved. doi:10.1016/j.lfs.2005.09.002

used by humans throughout time for recreational, spiritual, and therapeutic purposes (Lewin, 1924). Cannabis sativa (marijuana), Papaver somniferum (opium), Coffea arabica (caffeine), Nicotiana tabacum (tobacco), as well as other plants and plant-derived substances are widely used and abused at present. Investigation of psychoactive plants and their mechanisms of action have provided insight into the neurochemistry of many CNS diseases as well as the ‘‘chemistry of consciousness’’ (Lewin, 1924; Nichols, 2004). The observation that

K.A. O’Connor, B.L. Roth / Life Sciences 78 (2005) 506 – 511

507

serotonin and lysergic acid diethylamide, the analogue of ergot alkaloids produced by Claviceps purpurae, share structural and pharmacological properties led to the suggestion that biogenic amines, like serotonin, were involved in mental disorders such as schizophrenia (Gaddum and Hameed, 1954; Wooley and Shaw, 1954). Additionally, the active ingredient in Rauwolfa serpentina (resperine) has been shown to deplete biogenic amines and induce depression, therefore suggesting that a lack of serotonin and/or norepeinephrine may be the cause of this pathology (Vertulani and Sulser, 1975). Our basic understanding of mental illness as a neurochemical disease, as well as our ability to treat these disorders has been greatly enhanced through the study of psychoactive plants. Continued evaluation of the molecular site of action of psychoactive drugs will expand our list of validated targets for CNS drug discovery. Plants and plant-derived psychoactive compounds exert their actions via interaction with various signal transduction molecules, either a cell surface or intracellular recognition molecule (i.e. Freceptors_). It has recently been estimated that ¨20% of the recently completed human genome sequence is devoted to signal transduction, and specifically receptors (Lander et al., 2001; Venter et al., 2001). Sequencing the genome therefore, revealed a large number of potential targets for psychoactive drugs, and suggested the necessity of a comprehensive, systematic approach to identifying such targets. As summarized below, the development of high throughput screening (HTS) tools are necessary to screen psychoactive compounds against the receptorome. The receptorome Of the relatively large portion of the human genome dedicated to signal transduction, that portion dedicated to encoding ligand reception has been described as the ‘‘receptorome’’ (Kroeze et al., 2003) and encompasses more than 8% of the human genome (Roth et al., 2004). The receptorome is subdivided into multiple receptor superfamilies, the largest of which is that of the G-protein coupled receptors (GPCRs). The GPCR superfamily accounts for approximately 3.7% of the human genome: 735 – 802 open reading frames, of which ¨375 are neither olfactory nor taste receptors (Fredriksson et al., 2003; Kroeze et al., 2003; Roth et al., 2004). GPCRs are the most common molecular target for psychoactive drugs and
Table 1 Representative on-line resources for screening the receptorome Name NIH Blast Rasmol The Vaults of Erowid Entheogen dot The Lycaeum Botanical.com Multidisciplinary association for psychedelic studies Heffter Research Institute NIMH-PDSP database URL

pharmaceuticals (Hopkins and Groom, 2002; Kroeze et al., 2003; Lander et al., 2001; Vassilatis et al., 2003). Non-GPCR receptors represent at least 1.5% of the genome (Venter et al., 2001). Naturally occurring psychoactive compounds may also produce responses via ion channels and transporters, functioning as ‘‘receptors,’’ which represent an additional 3% of the genome (Roth et al., 2004; Venter et al., 2001). Receptoromics Virtual screening of the receptorome Multiple computational resources currently exist to assist in screening psychoactive compounds against the receptorome in a virtual (in silico) manner. Bioinformatic approaches such as BLAST have provided information on ligand:receptor interaction based on consensus domain profiling and hidden Markov models (HMM) (Gaulton and Attwood, 2003; Wise et al., 2004). Molecular modeling has also been used to virtually screen libraries of compounds for drug discovery efforts. For most GPCR, this classical computational approach has utilized homology modeling with rhodopsin (Ballesteros et al., 2001; Palczewski et al., 2000; Shapiro et al., 2002). This model has been quite successful in identifying molecular targets on the 5HT2A receptor, which interacts with many plant-derived hallucinogens (e.g., lysergic acid amide, psilocybin, mescaline) (Nichols, 2004; Roth et al., 2004). Molecular visualization software such as RasMol (http://www.umass.edu/microbio/ rasmol) is another useful tool for viewing structure:function relationships and may be accessed for free. Multiple on-line resources for identifying molecular targets for psychoactive botanicals also exist, and provide information for scientists and nonscientists alike (Table 1). Both The Vaults of Erowid (http://www.erowid.org) and The Lycaeum (http:// www.lycaeum.org) provide useful background on the chemistry, molecular targets, history and lore of psychoactive plants. Although these sites are not subject to peer review, they do provide useful information to the interested nonscientist such as anecdotal user reports, botanical information and links to additional articles. Databases, such as the National Institute of Mental Health’s Psychoactive Drug Screening Program (NIMH-PDSP) K i Database (http://kidb.cwru.edu), allow users to quickly identify

Type of information MM, S MM A,B,C,L,MT A,C,MT A,B,C,L,MT B L C,L,MT C,L,MT

http://www.ncbi.nlm.nih.gov/Education/BLASTinfo/information3.html http://www.umass.edu/microbio/rasmol/ http://www.erowid.org/ http://www.entheogen.com/ http://www.lycaeum.org/ http://www.botanical.com/ http://maps.org/ http://www.heffter.org/ http://kidb.cwru.edu/

A = anecdotal user reports; B = botanical information; C = chemistry; L = links to articles; MM = molecular modeling or visualization; MT = molecular target; S = sequencing information.

508

K.A. O’Connor, B.L. Roth / Life Sciences 78 (2005) 506 – 511

commonalities among drug –receptor interactions. This fully searchable database is updated daily and currently contains > 30,000 K i values from the literature and NIMH-PDSP screening initiative. The K i Database is a collection of organized tables and programs which include not only affinity values for specific compounds screened against a large group of receptors, but also the experimental conditions used to obtain these values (see Roth et al., 2004 for a complete description of the K i Database). Physical screening of the receptorome The identification of molecular targets of psychoactive compounds may also be studied using direct approaches based on ligand:receptor interactions and functional assays. Currently, the NIMH-PDSP is the single largest collection of receptors for which compounds may be physically screened. Traditionally, ligand binding experiments have been used to compete a test ligand and a receptor-specific high affinity radiolabeled ligand using either whole cell or membrane preparations (Pert and Snyder, 1973). This method may be utilized as a high throughput screen because (a) multiple receptors may be probed simultaneously against a given ligand, and (b) binding assays allow the use of frozen stocks of tissue or cellular membranes rather than live specimen. However, receptor binding has many disadvantages as well: (a) they do not identify the functional properties of ligands such as antagonist, agonist, partial or inverse agonist, (b) they do not properly assess allosteric ligands that may bind at sites other than the primary binding site. Furthermore, their reliance on high affinity, receptor-specific radiolabeled ligands reduces their use in identifying orphan receptors. Complimentary functional screens may be subsequently performed to either validate traditional binding assays or elucidate additional information. Due to the versatility in GPCR signal transduction, virtually every possible signal transduction pathway may be used to assess functional, high throughput screening of the receptorome. These assays take advantage of the well-characterized, diverse GPCR signaling cascades (Table 2). Receptor and/or G-protein activation, second-messenger production and even transcriptional activation may be assessed in a manner conducive to high throughput screening.
Table 2 HTS functional assays based on diverse GPCR signaling and regulation Function G-protein activation GTPase activity Second messenger Adenylyl cyclase activity production and Intracellular Ca2+ release activity Ion channel activity Transcriptional activation Desensitization Assay

HTS has also been applied to ion channel modulators and may be helpful for studying such naturally occurring compounds as Piper methysticum, Kava kava. This plant is though to act via blockade of voltage-gated sodium ion channels, and calcium ion channels. Additionally, kava has been used in the treatment of depression and anxiety (Martin et al., 2002; Singh and Singh, 2002). Screening of calcium channel activation by various agonists can be measured using calcium-sensitive dyes such as those used in the Fluorescent Imaging Plate Reader (FLIPR, Molecular Devices, Sunnyvale, CA). Recently, the Voltage/Ion Probe Reader (VIPR, Aurora Biosciences, Indianapolis, IN) has been modified to provide an automated screening for ion channel modulators (Falconer et al., 2002; Gonzalez and Maher, 2002). VIPR is a versatile assay in that it may be used to identify changes in Na+, K+, ClÀ, Ca++ and ligand-gated channels. It is therefore ideal for use as a high throughput screen (> 50,000 compounds/day) for either drug discovery of analysis of psychoactive compounds (Gonzalez and Maher, 2002). Molecular targets of naturally occurring psychoactive-compounds Salvia divinorum Salvinorin A, ‘‘the magic-mint’’ hallucinogen, is a naturally occurring and potent hallucinogen from the sage Saliva divinorum (Ortega et al., 1982; Valdes et al., 1984). Broad screening of the receptorome can be appreciated in the elucidation of the molecular target of salvinorin A. High throughput screening profiling determined that salvinorin A possessed high affinity and potency for the n opioid receptor (KOR) (Roth et al., 2002). Interestingly, the specific effects of salvinorin A were unique when compared to other hallucinogens since it did not have appreciable affinity for either A or y opioid, or 5-HT2A receptors (Chavkin et al., 2004; Roth et al., 2002). Recently in vivo studies have confirmed the hypothesis that salvinorin A exerts its effects via KOR. The nonselective opioid antagonist, naloxone, has been reported to block its effects in humans (Sheffler and Roth, 2003) and its effects in nonhuman primates were shown to be equivalent to standard KOR agonists (Butelman et al., 2004). These results may provide insight into the pathology and treatment of diseases

References (Asano et al., 1984; Sim et al., 1995) (Shirokova et al., 2005) (Coward et al., 1999; Wise et al., 2004) (Falconer et al., 2002; Gonzalez and Maher, 2002) (Burstein et al., 1995; Weiner et al., 2001) (Ferguson and Caron, 2004; Milligan, 2002; Parnot et al., 2002) (Janetopoulos et al., 2001)

Agonist-stimulated [35S]GTPgS binding Release of 32P-Pi from [g-32P]-GTP Stimulation/Inhibition of AC stimulated cAMP Fluorescent measures from PLCg driven IP3 production Fluorescent measures GPCR-mediated MAPK activation Cellular protein redistributions, enhanced cell surface expression of WT and constitutively active GPCRs Receptor dimerization and/or FRET, BRET G-protein dissociation

AC = adenylyl cyclase; BRET = bioluminescence resonance energy transfer; FRET = fluorescence resonance energy transfer; GTP = guanosine triphosphate; GTPase = guanosine triphosphatase; MAPK = mitogen activated protein kinase; PLC = phospholipase C.

K.A. O’Connor, B.L. Roth / Life Sciences 78 (2005) 506 – 511

509

associated with altered perception (e.g., schizophrenia, Alzheimer’s disease). Banisteriopsis caapi Ayahuasca is a South American psychotropic beverage which is made by boiling the bark of the Banisteriopsis caapi (Malpighiaceae) with one of various other psychotropic plants (Dobkin de Rios, 1984; Schultes and Hofmann, 1980). This psychotropic concoction has been used extensively in spiritual ceremonies and for medicinal purposes in the Upper Amazon River Basin (Schultes and Hofmann, 1982), and its use has recently spread to Europe and North America (Dobkin de Rios, 1996). The psychotropic plants added to the B. caapi mixture, such as Psychotria viridis (Rubiaceae), possess a potent hallucinogen N,N-dimethyltryptamine (DMT) while B. caapi contains beta-carbolines (e.g., harmine, harmaline, and tetrahydroharmine) which are highly active monoamine oxidase (MAO) inhibitors (Rivier and Lindgren, 1972; Schultes and Hofmann, 1980). This combination produced in ayahuasca renders DMT, normally inactive orally due to peripheral MAO activity (Suzuki et al., 1981), an efficacious psychoactive compound in the presence of beta-carbolines (Ott, 1999). DMT may then reach the brain where it acts as an agonist at 5-HT2A/2C receptors (McKenna et al., 1984; Pierce and Peroutka, 1989). Recent studies have revealed that ingestion of ayahuasca produced concentration-dependent specific inhibition of MAOA, in liver homogenates (Schwarz et al., 2003). Furthermore, harmine and harmaline also stimulate dopamine release in rat striatal slices (Schwarz et al., 2003). The recent finding that patients with Parkinson’s Disease and Parkinson’s-like symptoms report improvements following ingestion of ayahuasca (Sanchez-Ramos, 1991; Serrano-Duenas et al., 2001) may provide insight into additional natural-products-based treatments for this and other diseases. Conclusions While several psychoactive compounds such as C. sativa (THC), and N. tabacum (nicotine) have well-described mechanisms of action, the molecular targets of many hallucinogenic plants still elude scientists (e.g., Tabernanthe iboga and Myrstica fragens). When receptorome profiling has been done the results have been highly informative, however this has only been done for a few compounds (Roth et al., 2002; Rothman et al., 2003). Due to the recently sequenced genome and the rapid ‘‘deorphanizing’’ of many receptors, the potential molecular targets for these compounds are rapidly growing. A discovery-based effort seeking to elucidate these targets is likely to be highly useful for basic scientists as well as for drug development. Broad, unbiased receptorome screens, such as those described herein, may contribute to the detection of novel drug:receptor interactions, and ligand interactions with multiple receptors. Furthermore, receptorome-wide screening of psychoactive compounds may provide insight into novel

therapeutic compounds, possible side-effects of currently marketed pharmaceuticals and a more appropriate understanding of the neurochemical complexity of multifaceted diseases (e.g., schizophrenia, Alzheimer’s disease, depression, anxiety, drug abuse). References
Asano, T., Pedersen, S.E., Scott, C.W., Ross, E.M., 1984. Reconstitution of catecholamine-stimulated binding of guanosine 5V-O-(3-thiotriphosphate) to the stimulatory GTP-binding protein of adenylate cyclase. Biochemistry 23 (23), 5460 – 5467. Ballesteros, J.A., Shi, L., Javitch, J.A., 2001. Structural mimicry in G proteincoupled receptors: implications of the high-resolution structure of rhodopsin for structure-function analysis of rhodopsin-like receptors. Molecular Pharmacology 60 (1), 1 – 19. Burstein, E.S., Spalding, T.A., Hill-Eubanks, D., Brann, M.R., 1995. Structurefunction of muscarinic receptor coupling to G proteins. Random saturation mutagenesis identifies a critical determinant of receptor affinity for G proteins. Journal of Biological Chemistry 270 (7), 3141 – 3146. Butelman, E.R., Harris, T.J., Kreek, M.J., 2004. The plant-derived hallucinogen, salvinorin A, produces kappa-opioid agonist-like discriminative effects in rhesus monkeys. Psychopharmacology (Berlin) 172, 220 – 224. Chavkin, C., Sud, S., Jin, W., Stewart, J., Zjawiony, J.K., Siebert, D.J., Toth, B.A., Hufeisen, S.J., Roth, B.L., 2004. Salvinorin A, an active component of the hallucinogenic sage Salvia divinorum, is a highly efficacious kappa opioid receptor agonist: structural and functional considerations. Journal of Pharmacology and Experimental Therapeutics 308, 1197 – 1203. Coward, P., Chan, S.D., Wada, H.G., Humphries, G.M., Conklin, B.R., 1999. Chimeric G proteins allow a high-throughput signaling assay of Gi-coupled receptors. Analytical Biochemistry 270 (2), 242 – 248. Dobkin de Rios, M., 1984. Visionary Vine: Hallucinogenic Healing in the Peruvian Amazon. Waveland Press, Prospect Heights. Dobkin de Rios, M., 1996. Hallucinogens: Cross Cultural Perspectives. Waveland Press, Prospect Heights. Falconer, F., Smith, F., Surah-Narwal, S., Congrave, G., Liu, Z., Hayter, P., Ciaramella, G., Keighley, W., Haddock, P., Waldron, G., Sewing, A., 2002. High-throughput screening for ion channel modulators. Journal of Biomolecular Screening 7 (5), 460 – 465. Ferguson, S.S., Caron, M.G., 2004. Green fluorescent protein-tagged betaarrestin translocation as a measure of G protein-coupled receptor activation. Methods in Molecular Biology 237, 121 – 126. Fredriksson, R., Lagerstrom, M.C., Lundin, L.G., Schioth, H.B., 2003. The Gprotein coupled receptors in the human genome form five main families. Phylogenetic analysis, paralogon groups, and fingerprints. Molecular Pharmacology 63 (6), 1256 – 1272. Gaddum, J.H., Hameed, K.A., 1954. Drugs which antagonize 5-hydroxytryptamine. British Journal of Clinical Pharmacology 9, 240 – 248. Gaulton, A., Attwood, T.K., 2003. Bioinformatics approaches for the classification of G-protein coupled receptors. Current Opinion in Pharmacology 3 (2), 114 – 120. Gonzalez, J.E., Maher, M.P., 2002. Cellular fluorescent indicators and voltage/ion probe reader (VIPR) tools for ion channel and receptor drug discovery. Recept. Channels 8 (5 – 6), 283 – 295. Hopkins, A.L., Groom, C.R., 2002. The druggable genome. Natute Reviews. Drug Discovery 1 (9), 727 – 730. Janetopoulos, C., Jin, T., Devreotes, P., 2001. Receptor-mediated activation of heterotrimeric G-proteins in living cells. Science 291 (5512), 2408 – 2411. Kroeze, W.K., Sheffler, D.J., Roth, B.L., 2003. G-protein coupled receptors at a glance. Journal of Cell Science 116 (Pt 24), 4867 – 4869. Lander, E.S., Linton, L.M., Birren, B., Nusbaum, C., Zody, M.C., Baldwin, J., Devon, K., Dewar, K., Doyle, M., FitzHugh, W., Funke, R., Gage, D., Harris, K., Heaford, A., Howland, J., Kann, L., Lehoczky, J., LeVine, R., McEwan, P., McKernan, K., Meldrim, J., Mesirov, J.P., Miranda, C., Morris, W., Naylor, J., Raymond, C., Rosetti, M., Santos, R., Sheridan, A.,

510

K.A. O’Connor, B.L. Roth / Life Sciences 78 (2005) 506 – 511 Pert, C.B., Snyder, S.H., 1973. Opiate receptor: demonstration in nervous tissue. Science 179, 1011 – 1014. Pierce, P.A., Peroutka, S.J., 1989. Hallucinogenic drug interactions with neurotransmitter receptor binding sites in human cortex. Psychopharmacology (Berlin) 97, 118 – 122. Rivier, L., Lindgren, J.E., 1972. ‘‘Ayahuasca’’, the South American hallucinogenic drink: an ethnobotanical and chemical investigation. Economic Botany 26, 101 – 129. Roth, B.L., Baner, K., Westkaemper, R., Siebert, D., Rice, K.C., Steinberg, S., Ernsberger, P., Rothman, R.B., 2002. Salvinorin A: a potent naturally occurring nonnitrogenous kappa opioid selective agonist. Proceedings of the National Academy of Sciences of the United States of America 99 (18), 11934 – 11939. Roth, B.L., Lopez, E., Beischel, S., Westkaemper, R.B., Evans, J.M., 2004. Screening the receptorome to discover the molecular targets for plantderived psychoactive compounds: a novel approach for CNS drug discovery. Pharmacology & Therapeutics 102, 99 – 110. Rothman, R.B., Vu, N., Partilla, J.S., Roth, B.L., Hufeisen, S.J., Compton-Toth, B.A., Birkes, J., Young, R., Glennon, R.A., 2003. In vitro characterization of ephedrine-related stereoisomers at biogenic amine transporters and the receptorome reveals selective actions as norepinephrine transporter substrates. Journal of Pharmacology and Experimental Therapeutics 307 (1), 138 – 145. Sanchez-Ramos, J.R., 1991. Banisterine and Parkinson’s disease. Clinical Neuropharmacology 14 (5), 391 – 402. Schultes, R.E., Hofmann, A., 1980. The Botany and Chemistry of Hallucinogens. Charles C. Thomas, Springfield. Schultes, R.E., Hofmann, A., 1982. Plantas de los dioses: origenes del uso de los alucinogenos. Fondo de Cultura Economica, Mexico D.F. Schwarz, M.J., Houghton, P.J., Rose, S., Jenner, P., Lees, A.D., 2003. Activities of extract and constituents of Banisteriopsis caapi relevant to Parkinsonism. Pharmacology, Biochemistry and Behavior 75 (3), 627 – 633. Serrano-Duenas, M., Cardozo-Pelaez, F., Sanchez-Ramos, J.R., 2001. Scientific Review of Alternative Medicine 5, 127 – 132. Shapiro, D.A., Kristiansen, K., Weiner, D.M., Kroeze, W.K., Roth, B.L., 2002. Evidence for a model of agonist-induced activation of 5-hydroxytryptamine 2A serotonin receptors that involves the disruption of a strong ionic interaction between helices 3 and 6. Journal of Biological Chemistry 277 (13), 11441 – 11449. Sheffler, D.J., Roth, B.L., 2003. Salvinorin A: the ‘‘magic mint’’ hallucinogen finds a molecular target in the kappa opioid receptor. Trends in Pharmacological Sciences 24 (3), 107 – 109. Shirokova, E., Schmiedeberg, K., Bedner, P., Niessen, H., Willecke, K., Raguse, J.D., Meyerhof, W., Krautwurst, D., 2005. De-orphaning of olfactory receptors via cAMP/CNGA2 signaling in HeLa/Olf cells — G protein-dependent agonism and antagonism of odorants. Journal of Biological Chemistry 280 (12), 11807 – 11815 (Mar 25). Sim, L.J., Selley, D.E., Childers, S.R., 1995. In vitro autoradiography of receptor-activated G proteins in rat brain by agonist-stimulated guanylyl 5V[gamma-[35S]thio]-triphosphate binding. Proceedings of the National Academy of Sciences of the United States of America 92 (16), 7242 – 7246. Singh, Y.N., Singh, N.N., 2002. Therapeutic potential of kava in the treatment of anxiety disorders. CNS Drugs 16 (11), 731 – 743. Suzuki, O., Katsumata, Y., Oya, M., 1981. Characterization of eight biogenic indoleamines as substrates for type A and type B monoamine oxidase. Biochemical Pharmacology 30, 1353 – 1358. Valdes, J.L., Butler, W.M., Hatfield, G.M., Paul, A.G., Koreeda, M., 1984. Divinorin A: a psychotropic terpenoid and divinorin B from the hallucinogenic Mexican mint Salvia divinorum. Journal of Organic Chemistry 49, 4716 – 7720. Vassilatis, D.K., Hohmann, J.G., Zeng, H., Li, F., Ranchalis, J.E., Mortrud, M.T., Brown, A., Rodriguez, S.S., Weller, J.R., Wright, A.C., Bergmann, J.E., Gaitanaris, G.A., 2003. The G-protein coupled receptor repertoires of human and mouse. Proceedings of the National Academy of Sciences of the United States of America 100, 4903 – 4908. Venter, J.C., Adams, M.D., Myers, E.W., Li, P.W., Mural, R.J., Sutton, G.G., Smith, H.O., Yandell, M., Evans, C.A., Holt, R.A., Gocayne, J.D.,

Sougnez, C., Stange-Thomann, N., Stojanovic, N., Subramanian, A., Wyman, D., Rogers, J., Sulston, J., Ainscough, R., Beck, S., Bentley, D., Burton, J., Clee, C., Carter, N., Coulson, A., Deadman, R., Deloukas, P., Dunham, A., Dunham, I., Durbin, R., French, L., Grafham, D., Gregory, S., Hubbard, T., Humphray, S., Hunt, A., Jones, M., Lloyd, C., McMurray, A., Matthews, L., Mercer, S., Milne, S., Mullikin, J.C., Mungall, A., Plumb, R., Ross, M., Shownkeen, R., Sims, S., Waterston, R.H., Wilson, R.K., Hillier, L.W., McPherson, J.D., Marra, M.A., Mardis, E.R., Fulton, L.A., Chinwalla, A.T., Pepin, K.H., Gish, W.R., Chissoe, S.L., Wendl, M.C., Delehaunty, K.D., Miner, T.L., Delehaunty, A., Kramer, J.B., Cook, L.L., Fulton, R.S., Johnson, D.L., Minx, P.J., Clifton, S.W., Hawkins, T., Branscomb, E., Predki, P., Richardson, P., Wenning, S., Slezak, T., Doggett, N., Cheng, J.F., Olsen, A., Lucas, S., Elkin, C., Uberbacher, E., Frazier, M., Gibbs, R.A., Muzny, D.M., Scherer, S.E., Bouck, J.B., Sodergren, E.J., Worley, K.C., Rives, C.M., Gorrell, J.H., Metzker, M.L., Naylor, S.L., Kucherlapati, R.S., Nelson, D.L., Weinstock, G.M., Sakaki, Y., Fujiyama, A., Hattori, M., Yada, T., Toyoda, A., Itoh, T., Kawagoe, C., Watanabe, H., Totoki, Y., Taylor, T., Weissenbach, J., Heilig, R., Saurin, W., Artiguenave, F., Brottier, P., Bruls, T., Pelletier, E., Robert, C., Wincker, P., Smith, D.R., Doucette-Stamm, L., Rubenfield, M., Weinstock, K., Lee, H.M., Dubois, J., Rosenthal, A., Platzer, M., Nyakatura, G., Taudien, S., Rump, A., Yang, H., Yu, J., Wang, J., Huang, G., Gu, J., Hood, L., Rowen, L., Madan, A., Qin, S., Davis, R.W., Federspiel, N.A., Abola, A.P., Proctor, M.J., Myers, R.M., Schmutz, J., Dickson, M., Grimwood, J., Cox, D.R., Olson, M.V., Kaul, R., Shimizu, N., Kawasaki, K., Minoshima, S., Evans, G.A., Athanasiou, M., Schultz, R., Roe, B.A., Chen, F., Pan, H., Ramser, J., Lehrach, H., Reinhardt, R., McCombie, W.R., de la Bastide, M., Dedhia, N., Blocker, H., Hornischer, K., Nordsiek, G., Agarwala, R., Aravind, L., Bailey, J.A., Bateman, A., Batzoglou, S., Birney, E., Bork, P., Brown, D.G., Burge, C.B., Cerutti, L., Chen, H.C., Church, D., Clamp, M., Copley, R.R., Doerks, T., Eddy, S.R., Eichler, E.E., Furey, T.S., Galagan, J., Gilbert, J.G., Harmon, C., Hayashizaki, Y., Haussler, D., Hermjakob, H., Hokamp, K., Jang, W., Johnson, L.S., Jones, T.A., Kasif, S., Kaspryzk, A., Kennedy, S., Kent, W.J., Kitts, P., Koonin, E.V., Korf, I., Kulp, D., Lancet, D., Lowe, T.M., McLysaght, A., Mikkelsen, T., Moran, J.V., Mulder, N., Pollara, V.J., Ponting, C.P., Schuler, G., Schultz, J., Slater, G., Smit, A.F., Stupka, E., Szustakowski, J., Thierry-Mieg, D., Thierry-Mieg, J., Wagner, L., Wallis, J., Wheeler, R., Williams, A., Wolf, Y.I., Wolfe, K.H., Yang, S.P., Yeh, R.F., Collins, F., Guyer, M.S., Peterson, J., Felsenfeld, A., Wetterstrand, K.A., Patrinos, A., Morgan, M.J., Szustakowki, J., de Jong, P., Catanese, J.J., Osoegawa, K., Shizuya, H., Choi, S., Chen, Y.J., 2001. Initial sequencing and analysis of the human genome. Nature 409 (6822), 860 – 921. Lewin, L., 1924. Phantastica: Narcotic and Stimulating Drugs. Park Street Press, Rochester. Martin, H.B., McCallum, M., Stofer, W.D., Eichinger, M.R., 2002. Kavain attenuates vascular contractility through inhibition of calcium channels. Planta Medica 68 (9), 784 – 789. McKenna, D.J., Towers, G.H., Abbott, F.S., 1984. Monoamine oxidase inhibitors in South American hallucinogenic plants Part 2: constituents of orally active Myristicaceous hallucinogens. Journal of Ethnopharmacology 12 (2), 179 – 211. Milligan, G., 2002. Strategies to identify ligands for orphan G-protein coupled receptors. Biochemical Society Transactions 30 (4), 789 – 793. Nichols, D., 2004. Hallucinogens. Journal of Pharmacology and Experimental Therapeutics 101, 131 – 181. Ortega, A., Blount, J.F., Manchand, P.S., 1982. Salvinorin, a new transneoclerodane diterpene from Salvia divinorum (Labiatae). Journal of the Chemical Society. Perkin Transactions, 2505 – 2508. Ott, J., 1999. Pharmahuasca: human pharmacology of oral DMT plus harmine. Journal of Psychoactive Drugs 31, 171 – 177. Palczewski, K., Kumasaka, T., Hori, T., Behnke, C.A., Motoshima, H., Fox, B.A., Le Trong, I., Teller, D.C., Okada, T., Stenkamp, R.E., Yamamoto, M., Miyano, M., 2000. Crystal structure of rhodopsin: A G-protein coupled receptor. Science 289 (5480), 739 – 745. Parnot, C., Miserey-Lenkei, S., Bardin, S., Corvol, P., Clauser, E., 2002. Lessons from constitutively active mutants of G-protein coupled receptors. Trends in Endocrinology and Metabolism 13 (8), 336 – 343.

K.A. O’Connor, B.L. Roth / Life Sciences 78 (2005) 506 – 511 Amanatides, P., Ballew, R.M., Huson, D.H., Wortman, J.R., Zhang, Q., Kodira, C.D., Zheng, X.H., Chen, L., Skupski, M., Subramanian, G., Thomas, P.D., Zhang, J., Gabor Miklos, G.L., Nelson, C., Broder, S., Clark, A.G., Nadeau, J., McKusick, V.A., Zinder, N., Levine, A.J., Roberts, R.J., Simon, M., Slayman, C., Hunkapiller, M., Bolanos, R., Delcher, A., Dew, I., Fasulo, D., Flanigan, M., Florea, L., Halpern, A., Hannenhalli, S., Kravitz, S., Levy, S., Mobarry, C., Reinert, K., Remington, K., AbuThreideh, J., Beasley, E., Biddick, K., Bonazzi, V., Brandon, R., Cargill, M., Chandramouliswaran, I., Charlab, R., Chaturvedi, K., Deng, Z., Di Francesco, V., Dunn, P., Eilbeck, K., Evangelista, C., Gabrielian, A.E., Gan, W., Ge, W., Gong, F., Gu, Z., Guan, P., Heiman, T.J., Higgins, M.E., Ji, R.R., Ke, Z., Ketchum, K.A., Lai, Z., Lei, Y., Li, Z., Li, J., Liang, Y., Lin, X., Lu, F., Merkulov, G.V., Milshina, N., Moore, H.M., Naik, A.K., Narayan, V.A., Neelam, B., Nusskern, D., Rusch, D.B., Salzberg, S., Shao, W., Shue, B., Sun, J., Wang, Z., Wang, A., Wang, X., Wang, J., Wei, M., Wides, R., Xiao, C., Yan, C., Yao, A., Ye, J., Zhan, M., Zhang, W., Zhang, H., Zhao, Q., Zheng, L., Zhong, F., Zhong, W., Zhu, S., Zhao, S., Gilbert, D., Baumhueter, S., Spier, G., Carter, C., Cravchik, A., Woodage, T., Ali, F., An, H., Awe, A., Baldwin, D., Baden, H., Barnstead, M., Barrow, I., Beeson, K., Busam, D., Carver, A., Center, A., Cheng, M.L., Curry, L., Danaher, S., Davenport, L., Desilets, R., Dietz, S., Dodson, K., Doup, L., Ferriera, S., Garg, N., Gluecksmann, A., Hart, B., Haynes, J., Haynes, C., Heiner, C., Hladun, S., Hostin, D., Houck, J., Howland, T., Ibegwam, C., Johnson, J., Kalush, F., Kline, L., Koduru, S., Love, A., Mann, F., May, D., McCawley, S., McIntosh, T., McMullen, I., Moy, M., Moy, L., Murphy, B., Nelson, K., Pfannkoch, C., Pratts, E., Puri, V., Qureshi, H., Reardon, M., Rodriguez, R., Rogers, Y.H., Romblad, D., Ruhfel, B., Scott, R., Sitter, C., Smallwood, M., Stewart, E., Strong, R., Suh, E., Thomas, R., Tint, N.N., Tse, S., Vech, C., Wang, G., Wetter, J., Williams, S., Williams, M., Windsor, S., Winn-Deen, E., Wolfe, K., Zaveri, J., Zaveri, K., Abril, J.F., Guigo, R.,

511

Campbell, M.J., Sjolander, K.V., Karlak, B., Kejariwal, A., Mi, H., Lazareva, B., Hatton, T., Narechania, A., Diemer, K., Muruganujan, A., Guo, N., Sato, S., Bafna, V., Istrail, S., Lippert, R., Schwartz, R., Walenz, B., Yooseph, S., Allen, D., Basu, A., Baxendale, J., Blick, L., Caminha, M., Carnes-Stine, J., Caulk, P., Chiang, Y.H., Coyne, M., Dahlke, C., Mays, A., Dombroski, M., Donnelly, M., Ely, D., Esparham, S., Fosler, C., Gire, H., Glanowski, S., Glasser, K., Glodek, A., Gorokhov, M., Graham, K., Gropman, B., Harris, M., Heil, J., Henderson, S., Hoover, J., Jennings, D., Jordan, C., Jordan, J., Kasha, J., Kagan, L., Kraft, C., Levitsky, A., Lewis, M., Liu, X., Lopez, J., Ma, D., Majoros, W., McDaniel, J., Murphy, S., Newman, M., Nguyen, T., Nguyen, N., Nodell, M., Pan, S., Peck, J., Peterson, M., Rowe, W., Sanders, R., Scott, J., Simpson, M., Smith, T., Sprague, A., Stockwell, T., Turner, R., Venter, E., Wang, M., Wen, M., Wu, D., Wu, M., Xia, A., Zandieh, A., Zhu, X., 2001. The sequence of the human genome. Science 291 (5507), 1304 – 1351. Vertulani, J., Sulser, F., 1975. Action of various antidepressant treatments reduces reactivity of noradrenergic cyclic AMP-generating system in limbic forebrain. Nature 257, 495 – 496. Weiner, D.M., Burstein, E.S., Nash, N., Croston, G.E., Currier, E.A., Vanover, K.E., Harvey, S.C., Donohue, E., Hansen, H.C., Andersson, C.M., Spalding, T.A., Gibson, D.F., Krebs-Thomson, K., Powell, S.B., Geyer, M.A., Hacksell, U., Brann, M.R., 2001. 5-hydroxytryptamine2A receptor inverse agonists as antipsychotics. Journal of Pharmacology and Experimental Therapeutics 299 (1), 268 – 276. Wise, A., Jupe, S.C., Rees, S., 2004. The identification of ligands at orphan Gprotein coupled receptors. Annual Review of Pharmacology and Toxicology 44, 43 – 66. Wooley, D.W., Shaw, E., 1954. A biochemical and pharmacological suggestion about certain mental disorders. Proceedings of the National Academy of Sciences of the United States of America 40, 228 – 231.

Sign up to vote on this title
UsefulNot useful