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2011 International Conference on Bioscience, Biochemistry and Bioinformatics IPCBEE vol.5 (2011) © (2011) IACSIT Press, Singapore Virtual Screening of Active Compounds of Valeriana wallichii, Acorus Calamus and Asparagus Racemosus with Schizophrenic Proteins COMT & GRM3 Preenon Bagchi Waheeta Hopper Research Scholar, Dept. of Bioinformatics, School of Bioenginering, SRM University, Kattankulathur, Tamil Nadu, India e.mail: [email protected] Professor & Head, Dept. of Bioinformatics, School of Bioenginering, SRM University, Kattankulathur, Tamil Nadu, India e.mail: [email protected] involved with personality, planning, inhibition of behaviors, abstract thinking, emotion, and working (short-term) memory. To function efficiently, the prefrontal cortex requires signalling by neurotransmitters such as dopamine and norepinephrine. The characteristic signs and symptoms of 22q11.2 deletion syndrome result from a deletion of a small piece of chromosome 22. A loss of one copy of the COMT gene leads to abnormal regulation of catechol-omethyltransferase levels in the brain. Researchers believe that changes involving this enzyme in the prefrontal cortex may help to explain the increased risk of behavioral problems and mental illness associated with 22q11.2 deletion syndrome. People with 22q11.2 deletion syndrome are more likely to develop schizophrenia, depression, anxiety, and bipolar disorder than people without the condition [3, 4]. Abnormally high dopaminergic transmission has been linked to psychosis and schizophrenia. Increased dopaminergic functional activity, specifically in the mesolimbic pathway, is found in schizophrenic individuals. Anti-psychotic medications act largely as dopamine antagonists, inhibiting dopamine at the receptor level, and thereby blocking the effects of the neurochemical in a dose-dependent manner. Inhibitors of alternative metabolic route for dopamine by catechol-o-methyl transferase are also used [5]. GRM3: Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system. Imbalances in glutamatergic function have been implicated in neuronal death following ischemia, in hypoglycemia or anoxia, in epilepsy, and in neurodegenerative disorders. Glutamate actions are mediated by glutamate receptors which fall into 2 distinct classes: ionotropic and metabotropic receptors. Ionotropic glutamate receptors are ligand-gated ion channels whose responses to selective agonists define the N-methyl-D-aspartate, alpha-amino-3hydroxy-5-methyl-isoxasole-4-propionate, and kainate subtypes. The metabotropic glutamate receptors, which are coupled through GTP-binding proteins to second-messenger pathways, can be involved in the stimulation of phospholipase C, the presynaptic inhibition of glutamate release, the closure of cation channels in retinal on bipolar cells, and the modulation of adenylate cyclase. The eight distinct metabotropic glutamate receptors that have been identified can be classified into three groups according to their sequence similarities, pharmacologic properties, and Abstract—Gene mutation in COMT and GRM3 are one of the causes for Schizophrenia. Their amino acid sequences are retrieved; their 3D structures were determined by homology modelling. The 3D structures of the compounds of Valeriana wallichii, Acorus calamus and Asparagus racemosus were sketched using Chemsketch & converted to 3D. These compounds were virtually screened with COMT and GRM3 proteins. COMT protein had good interaction with baldrinal from V. wallichii, calamusenone from A. calamus and racemosol from A. racemosus. GRM3 protein had good interaction with homobaldrinal from V. wallichii, galangin from A. calamus and racemosol from A. racemosus. Keywords-Schizophrenia, Homology Modelling, Valeriana wallichii, Acorus calamus, Asparagus racemosus, metabotropic glutamate receptor, Virtual Screening. I. INTRODUCTION Schizophrenia is a severe mental disorder characterized by two kinds of symptoms; positive psychotic symptoms thought disorder, hallucinations, delusions, and paranoia and negative symptoms – impairment in emotional range, energy, and enjoyment of activities. Schizophrenia is a particular form of psychosis, a term encompassing several severe mental disorders that result in the loss of contact with reality along with major personality derangements. Presently researchers suspect a genetical cause for schizophrenia along with environmental basis. Psychosocial factors play an important role in the formation & development of this disorder. Individual as well as family counseling is necessary to help patients to lead a normal social life. Cognitivebehavioral approach along with proper medication can pave a path for curing this disorder. The cause of Schizophrenia is multi-gene mutation (including environmental factors) [1, 2]. In this work COMT and GRM3 genes are virtually screened with the compounds of Valeriana wallichii, Acorus calamus and Asparagus racemosus. COMT: Catechol-O-methyltransferase catalyzes the transfer of a methyl group from s-adenosylmethionine to catecholamines, including the neurotransmitters dopamine, epinephrine, and norepinephrine. Catechol-omethyltransferase is particularly important in the prefrontal cortex of the brain, which organizes and coordinates information from other parts of the brain. This region is 15 preferred signal transduction mechanism. The human genes for mGlu2 and mGlu3 receptors have been localized to chromosomes 3 (p21.1) and 7 (7q21.1-q21.2) respectively. The main glutamate transporter GLT-1 is responsible for clearing synaptically released glutamate from the extracellular space and contributes to the shaping of glutamatergic transmission [6, 7]. Valeriana wallichii: Valeriana wallichii commonly known as Indian valerian is one of the important plant species of commerce. It is native to India (Himalayas). It grows wild in the temperate Himalaya at an altitude of 15003000 m and is an ingredient of Indian systems of herbal medicine. It is used in India for its benefits in calming down the nervous system. It relieves stress and anxiety and also fights depression. Valeriana is nature's best stress busting herb. Valeriana continues to be a safe sedative/hypnotic choice for patients with mild to moderate insomnia. The compounds identified from the plant are acetoxy valerenic acid, acevaltrate, baldrinal, bornyl acetate, bornyl isovalerate, fenchene, β-sitosterol, calarene, homobaldrinal, isovaltrate, valeranone, valerenal, valerenic acid, valepotriate, valtrate, valtroxal and xanthorrhizol (Fig. 1) [8]. Acorus calamus: Acorus calamus or sweet flag has been long known for its medicinal value and is cultivated in Asia for this reason. The rhizome contains aromatic oil that has been used medicinally since ancient times. The rhizome possesses anti-spasmodic, carminative and anthelmintic properties and is also used for the treatment of epilepsy, mental ailments, chronic diarrhea, dysentery, bronchial catarrh, intermittent fevers and glandular and abdominal tumors. The compounds identified from the plant are 1α, 2β, 3γ, 19α-tetrahydroxyurs-12en-28-oicacid-28-O{-β-Dglucopyranosyl (1→2)} β- D- galactopyranoside, 2,3dihydro-4,5,7-trimethoxy-1ethyl-2-methyl-3-(2,4,5trimethoxy phenyl)indene, 2,4,5-trimethoxy benzaldehyde, 2,6-diepishyobunone, 3β, 22α, 24, 29-tetrahydroxyolean-12en-3-O-{-β-D-arabinosyl(1→3)}-β-D-arabinopyranoside, 4, 5, 8-trimethoxyxanthone-2-O-β-D-glucopyranosyl(1→2)-Oβ-D-galactopyranoside, acoradin, acoragermacrone, acoramone(1,2,4 –trimethoxy-5(2-propanoyl) benzene, β-sitosterol, Calamusenone, cis-asarone(cis-1,2,4 – trimethoxy-5(2propenyl) benzene, galangin, γ-cis-asarone(cis-1,2,4 –trimethoxy-5(2- propenyl) benzene, isoeugenol methyl ether, isocalamendiol limonene, preisocalamendiol, shyobunone thujane and Z3-(2,4,5-trimethoxy phenyl)-2-propenal. (Fig. 2) [9]. Asparagus racemosus: Asparagus racemosus has been used in India for thousands of years for its therapeutic and tonic properties. Due to its multiple uses, the demand for Asparagus is constantly on the rise. The plant has been shown to aid in the treatment of neurodegenerative disorders and in alcohol abstinence-induced withdrawal symptoms. The genus Asparagus includes about 300 species around the world. The genus is considered to be medicinally important because of the presence of steroidal saponins and sapogenins in various parts of the plant. Out of the 22 species of Asparagus recorded in India, Asparagus racemosus is the one most commonly used in traditional medicine. The compounds identified from the plant are asparagamine, racemosol, sarsasapogenin and shatavarin (Fig 3) [10]. II. METHODOLOGY The COMT sequence with accession number NP_009294.1 was taken the National Center for Biotechnology Information (NCBI) and using BLAST search engine against Protein Data Bank (PDB) the following templates were selected and their crystal structures were downloaded from PDB. • 3BWMA: Crystal Structure Of Human Catechol OMethyltransferase with Bound Sam And Dnc. from Homo Sapiens [identity-96%] • 2ZLBA: Crystal Structure Of Apo Form Of Rat Catechol-O-Methyltransferase [identity-61%] • 1JR4A: Catechol O-Methyltransferase BisubstrateInhibitor Complex from Rattus Norvegicus [identity-98%] The COMT sequence is aligned with 3BWMA, 2ZLBA and 1JR4A. _aln.pos 3BWMA 2ZLBA 1JR4A COMT _consrvd 10 20 30 40 50 60 -GDTKEQRILNHVLQHAEPGNAQSVLEAIDTYCEQKEWAMNVGDKKGKIVDAVIQEHQPSVLLELGAY --DTKEQRILRYVQQNAKPGDPQSVLEAIDTYCTQKEWAMNVGDAKGQIMDAVIREYSPSLVLELGAY --DTKEQRILRYVQQNAKPGDPQSVLEAIDTYCTQKEWAMNVGDAKGQIMDAVIREYSPSLVLELGAY MGDTKEQRILNHVLQHAEPGNAQSVLEAIDTYCEQKEWAMNVGDKKGKIVDAVIQEHQPSVLLELGAY ******** * * * ** *********** ********** ** * **** * ** ****** _aln.p 3BWMA 2ZLBA 1JR4A COMT _consrvd 70 80 90 100 110 120 130 CGYSAVRMARLLSPGARLITIEINPDCAAITQRMVDFAGVKDKVTLVVGASQDIIPQLKKKYDVDTLD CGYSAVRMARLLQPGARLLTMEMNPDYAAITQQMLNFAGLQDKVTILNGASQDLIPQLKKKYDVDTLD CGYSAVRMARLLQPGARLLTMEMNPDYAAITQQMLNFAGLQDKVTILNGASQDLIPQLKKKYDVDTLD CGYSAVRMARLLSPGARLITIEINPDCAAITQRMVDFAGVKDKVTLVVGASQDIIPQLKKKYDVDTLD ************ ***** * * *** ***** * *** **** ***** ************** _aln.pos 3BWMA 2ZLBA 1JR4A COMT _consrvd 140 150 160 170 180 190 200 MVFLDHWKDRYLPDTLLLEECGLLRKGTVLLADNVICPGAPDFLAHVRGSSCFECTHYQSFLEYREVV MVFLDHWKDRYLPDTLLLEKCGLLRKGTVLLADNVIVPGTPDFLAYVRGSSSFECTHYSSYLEYMKVV MVFLDHWKDRYLPDTLLLEKCGLLRKGTVLLADNVIVPGTPDFLAYVRGSSSFECTHYSSYLEYMKVV MVFLDHWKDRYLPDTLLLEECGLLRKGTVLLADNVICPGAPDFLAHVRGSSCFECTHYQSFLEYREVV ******************* **************** ** ***** ***** ****** * *** ** _aln.pos 3BWMA 2ZLBA 1JR4A COMT _consrvd 210 220 DGLEKAIYKGP-----DGLEKAIYQG------DGLEKAIYQG------DGLEKAIYKGPGSEAGP ******** * Based on this data the 3D structure of COMT protein was generated by modeler. Also, GRM3 sequence with accession number AAA52568 was taken from NCBI. Using BLAST search engine against PDB , the following templates were selected: • 3KG2A: Ampa subtype ionotropic Glutamate Receptor in complex with competitive antagonist from Rattus Norvegicus [identity-89%] • 3H6GA: Crystal structure of the Glur6 amino terminal domain dimer assembly from Rattus Norvegicus [identity-43%] • 1YAEA: Structure of The kainate receptor subunit Glur6 agonist binding domain complexed with domoic acid from Rattus Norvegicus [identity-43%] The crystal structure of the above templates were downloaded from PDB. _aln.pos 3KG2A 3H6GA 1YAEA 16 10 20 30 40 50 60 -------------------------------------------NSIQIGGLFPRGADQEYSAFRVGMV ------------------------------------------------------------------------------------------------------PANITDSL------------------------- The 3d structures of the active components of Valeriana wallichii, Acorus calamus and Asparagus racemosus were drawn using chemsketch and saved as *.mol file. Virtual Screening of the compounds of Valeriana wallichii, Acorus calamus and Asparagus racemosus was done with COMT & GRM3 proteins using Maestro9.1 software. The proteins were prepared using Protein Preparation Wizard of Maestro9.1 software, optimized and minimized. The receptor grid was set up and active site residues for the proteins were assigned by using the Receptor Grid Generation panel. The grid region assigned for COMT protein was TRP38, MET40, ASP141, TRP143, LYS144, ASN170, VAL173, GLU199, MET201 and VAL203 [11]. The grid region assigned for GRM 3 protein was 74, 108, 110, 165, 188, 186, 191, 233, 236, 238, 242, 260, 318, 323 and 409 [12]. The ligands (compounds of the plants V. wallichii, A. calamus and A. racemosus) were converted to a single file. Using LigPrep, energy minimization was done. The Virtual Screening was carried out using the receptor and ligand as mentioned above. GRM3 MEHGTLLAQPGLWTRDTSWALLYFLCYILPQTAPQVLRIGGIFETVENEPVNVEELAFKFAVTSINRN _consrvd _aln.p 3KG2A 3H6GA 1YAEA GRM3 _consrvd 70 80 90 100 110 120 130 QFSTSEFRLTPHIDNLEVANSFAVTNAFCSQFSRGVYAIFGFYDKKSVNTITSFCGTLHVSFITPSFP --------------------------------------------------------------------------------------------------------------------------------------RTLMPNTTLTYDIQRINLFDSFEASRRACDQLALGVAALFGPSHSSSVSAVQSICNALEVPHIQTRWK _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 140 150 160 170 180 190 200 TDG------THPFVIQMRPDLKGALLSLIEYYQWDKFAYLYDSDRGLSTLQAVLDSAAEKKWQVTAIN --------------------------------------------------------------------------------------------------------------------------------------HPSVDNKDLFYINLYPDYAAISRAILDLVLYYNWKTVTVVYEDSTGLIRLQELIKAP-----SRYNIK _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 210 220 230 240 250 260 270 VGNINNDKKDETYRSLFQDLELKKERRVILDCERDKVNDIVDQVITIGKHVKGYHYIIANLGFTDGDL --------------------------------------------------------------------------------------------------------------------------------------IKIRQLPSGNKDAKPLLKEMKKGKEFYVIFDCSHETAAEILKQILFMGMMTEYYHYFFTTLDLFALDL _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 280 290 300 310 320 330 340 LKIQFGGAEVSGFQIVDYDDSLVSKFIERWSTLEEKEYPGAHTATIKYTSALTYDAVQVMTEAFRNLR --------------------------------------------------------------------------------------------------------------------------------------ELYRYSGVNMTGFRLLNIDNPHVSSIIEKWSMERLQAPPRPETGLLDGMMTTEAALMYDAVYMVAIAS _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 350 360 370 380 390 400 KQRIEISRRGNAGDCLANPAVPWGQGVEIERALKQVQVEGLSGNIKFDQ-NGKRINYTINIMELKTNG --------------------------------------------------------------------------------------------------------------------------------------HRASQLTVSSLQCHR----HKPWRLGPRFMNLIKEARWDGLTGHITFNKTNGLRKDFDLDIISLKEEG _aln.p 410 420 430 440 450 460 470 3KG2A PRKIGYWSEVDKMVLTEDDTSGLEQKTVVVTT----------------------ILESPYVMMKANHA 3H6GA ------------------------THVLRFGGIFEYVES---------GPMGAEELAFRFAVNT---1YAEA -----------------------SNRSLIVTT----------------------ILEEPYVLFKKSDK GRM3 TEKAAGEVSKHLYKVWKKIGIWNSNSGLNMTDSNKDKSSNITDSLANRTLIVTTILEEPYVMYRKSDK _consrvd * _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 480 490 500 510 520 530 540 ALAGNERYEGYCVDLAAEIAKHCGFKYKLTIVGDGKYGARDADTKIWNGMVGELVY--GKADIAIAPL ----INRNR----TLLPN-------TTLTYDTQK----INLYDSFEA---SKKACDQLSLGVAAIFGP PLYGNDRFEGYCIDLLRELSTILGFTYEIRLVEDGKYGAQDDVNGQWNGMVRELID--HKADLAVAPL PLYGNDRFEGYCLDLLKELSNILGFIYDVKLVPDGKYGAQN-DKGEWNGMVKELID--HRADLAVAPL * * * _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 550 560 570 580 590 600 610 TI---TLVREEVID--------------------FSKPFMSLGI--SIMIKKPQKSKPGVFSFLDPLA SHSSSANAVQSICNALGVPHIQTRWKHQVSDNKDSFYVSLYPDFSSLS-------------------AI---TYVREKVID--------------------FSKPFMTLGI--SILYRK---------------TI---TYVREKVID---F-----------------SKPFMTLGI--SILYRKPNGTNPGVFSFLNPLS _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 620 630 640 650 660 670 680 YEIWMCIVFAYIGVSVVLFLVST------------N--------EFGIFNSLWFSLGAFMQPRS-----------------------------------------------------------------------------------------------------------------------------------------PDIWMYVLLACLGVSCVLFVIARFTPYEWYNPHPCNPDSDVVENNFTLLNSFWFGVGALMQQGSELMP _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 690 700 710 720 730 740 --LSGRIVGGVWWFFTLIIISSYTANLAAFLTVERMVSPIESAE----------DLSKQTEIAYGTLD ----------------------------------------RAILDLVQFFKWKTVTVVY-----------------------------------------------IDSAD----------DLAKQTKIEYGAVE KALSTRIVGGIWWFFTLIIISSYTANLAAFLTVERMESPIDSAD----------DLAKQTKIEYGAVR III. Amino acid sequence of COMT and GRM3 were retrieved from NCBI’s database. Homology modelling of COMT was carried out using the software Modeller9v8 with the templates 3BWMA, 2ZLBA and 1JR4A (structure retrieved from RCSB’s pdb database) and five models were generated. Rampage Ramachandran Plot server was used to generate the values of COMT protein obtained in favoured, allowed and outlier region (Table I, Fig. 4). As per Ramachandran Plot, either Model 2 or Model 5 was selected as the best protein since both are having same and maximum number of the residues in favoured region (also same number of residues in allowed region and outlier region) (Fig. 5). Homology modelling of GRM3 protein was carried out using the software Modeller9v8. The templates used for GRM3 mutation protein were 3KG2A, 3H6GA, 1YAEA and five models (3D structures) of the GRM3 mutation protein were generated. Rampage Ramachandran Plot server generated the values of GRM3 protein obtained in favoured, allowed and outlier region (Table 2, Fig. 6). As per the Ramachandran Plot, Model 4 was selected as the best protein since it is having least residues in the outlier region (Fig. 7): The COMT protein (model #2 or 5) was virtually screened with the compounds of V. wallichii. Glide XP visualizer showed COMT’ LYS144 interacting with baldrinal from V. wallichii. The glide score was -6.09 (Fig. 8). COMT protein (model #2 or 5) was screened with the compounds of A. calamus. Glide XP visualizer showed COMT’ LYS144 interacts with calamusenone from A. calamus. The glide score was -5.23 (Fig. 9). _aln.p 750 760 770 780 790 800 810 3KG2A SGSTKEFFRRSKIAVFD--------------KMWTYMRSAEPS-------------------VFVRT3H6GA -------DDSTGLIRLQELIKAPSRYNLRLKIR--QLPADTKDAKPLLKEMKRGKEFHVIFDCSHEMA 1YAEA DGATMTFFKKSKISTYD--------------KMWAFMSSRRQS-------------------VLVKSGRM3 DGSTMTFFKKSKISTYE--------------KMWAFMSSRQQT-------------------ALVR-_consrvd _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 820 830 840 850 860 870 880 ----------------------TAEGVARVRKSK-----GKYAYLLESTMNEYIEQR----------AGILKQALAMGMMTEYYHYIFTTLDLFALDVEPYRYSGVNMTGF------RILNTENTQVSSIIEKWS ----------------------NEEGIQRVLTS-------DYAFLMESTTIEFVTQR------------N-------------------SDEGIQRVLTT-------DYALLMESTSIEYVTQRNCNLTQIGGL- _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 890 900 910 920 930 940 950 -----------------------------------------------KPCDTMKVGGNLDSKGYGIAT MEKPDSGLLDGFMTTDAALMYDAVHVVSVAVQQFPQMTVSSLQCNRH-------------------------------------------------------------------N-CNLTQIGGLIDSKGYGVGT -----------------IDSKGYGVGTPIGSPYRDKITIAILQLQEEGKLHMMKEKWWRGNGCPEEDN _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 960 970 980 990 1000 1010 1020 PKGSSLGTPVNLAVLKLSEQGLLDKLKNKWWYDKGECGA-------------------------KDSG -KPWRFGTRFMSLIKEAH--------WEGLTGR------ITFNKTNGLRTDFDLDVISLKEEGLEKIG PMGSPYRDKITIAILQLQEEGKLHMMKEKWWRG---NGC----------------------------KEASALGVENIGGIFIVLAAGLVLSVFVAIGEFIYKSRKNNDIEQAFCFFYGLQCKQTHPTNSTSGTT _aln.pos 3KG2A 3H6GA 1YAEA GRM3 _consrvd 1030 1040 1050 1060 S-------KEKTSALSLSNVAGVFYILVGGLGLAMLVALIEFCYK TWDPASGLNMTE----------------------------------------------------------------------------LSTDLECGKLIREERGIRKQSSVHTV------------------- RESULTS & DISCUSSION Based on this data, the 3D structure of GRM3 protein was generated by modeler. 17 COMT protein (model #2 or 5) was virtually screened with the compounds of A. racemosus. Glide XP visualizer showed COMT’ GLU90 interacting with racemosol from A. racemosus. The glide score obtained was –6.25 (Fig. 10). The GRM3 protein (model #4) was virtually screened with the compounds of V. wallichii. Glide XP visualize showed GRM3’s LYS59 interacting with homobaldrine of V. wallichii. The glide score was -6.59 (Fig. 11). The GRM3 protein (model #4) was virtually screened with the compounds of Acorus calamus. Glide XP visualize showed GRM3’ TYR154 & ASP230 interacting with galangin from Acorus calamus. The glide score was -7.77 (Fig. 12). The GRM3 protein (model #4) was virtually screened with the compounds of A.racemosus. Glide XP visualizer showed GRM3’s THR133 & ASP230 interacting with racemosol from A. racemosus. The glide score obtained was -8.37 (Fig. 13). IV. [11] Lee JY and Kim Y, “Comparative Homology Modeling and Ligand Docking Study of Human Catechol-O_Methyltransferase for Antiparkinson Drug Design,” Bull. Korean Chem. Soc. (2005) 26(11):1695-1700. [12] Tsuchiya D, Kunishima N, Kamiya N, Jingami H and Morikawa K, “Structural views of the ligand-binding cores of a metabotropic glutamate receptor complexed with an antagonist and both glutamate and Gd3+,” PNAS (2002) 99(5):2660-2665. TABLE I. VALUES OF COMT PROTEIN OBTAINED IN FAVOURED, ALLOWED AND OUTLIER REGION USING RAMPAGE RAMACHANDRAN PLOT SERVER Number of residues in favoured region (~98.0% expected) 213 (97.3%) Number of residues in allowed region (~2.0% expected) 5 (2.3%) Number of residues in outlier region Model 2 215 (98.2%) 3 (1.4%) 1 (0.5%) Model 3 213 (97.3%) 5 (2.3%) 1 (0.5%) Model 4 212 (96.8%) 6 (2.7%) 1 (0.5%) Model 5 215 (98.2%) 3 (1.4%) 1 (0.5%) Model 1 CONCLUSION COMP protein showed good interaction with baldrinal from Valeriana wallichii, calamusenone from Acorus calamus and racemosol from Asparagus racemosus. GRM3 protein showed good interaction with homobaldrinal from Valeriana wallichii, galangin from Acorus calamus and racemosol from Asparagus racemosus. These compounds, baldrinal, calamusenone, homobaldrine, galangin and racemosol, could be tested invitro and in-vivo for their efficacy in treating the disorder. Selected 1 (0.5%) selected selected REFERENCES TABLE II. VALUES OF COMT PROTEIN OBTAINED IN FAVOURED, ALLOWED AND OUTLIER REGION USING RAMPAGE RAMACHANDRAN PLOT SERVER. [1] Prasad S, Semwal P, Deshpande S, Bhatia T, Nimgaonkar V L and Thelma B K, “Molecular genetics of Schizophrenia: past, present and future,” J. Biosci. (2002) 27(1):35-52. [2] Kirov G, O'Donovan MC and Owen MJ, “Finding Schizophrenia Genes,” J Clin Invest. (2005) 115(6):1440-1448 [3] Baker K, Baldeweg T, Sivagnanasundaram S, Scambler P, Skuse D., “COMT Val108/158 Met modifies mismatch negativity and cognitive function in 22q11 deletion syndrome,” Biol Psychiatry. 2005 Jul 1;58(1):23-31. [4] Bearden CE, Jawad AF, Lynch DR, Monterossso JR, Sokol S, McDonald-McGinn DM, Saitta SC, Harris SE, Moss E, Wang PP, Zackai E, Emanuel BS, Simon TJ., “Effects of COMT genotype on behavioral symptomatology in the 22q11.2 Deletion Syndrome,” Neuropsychol Dev Cogn C Child Neuropsychol. 2005 Feb;11(1):10917. [5] Jones H. M. and Pilowsky L. S., “Dopamine and antipsychotic drug action revisited,” Br. J. Psychiatry, (2002) 181: 271-275 [6] Palmada M and Centelles JJ, “Excitatory Amino Acid Neurotransmission. Pathways For Metabolism, Storage And Reuptake Of Glutamate In Brain” (1998) Front Biosci., 3, 701-718 [7] Michael FE, et. al., “Variation in GRM3 affects cognition, prefrontal glutamate, and risk for schizophrenia” (2004) PNAS. 101(34): 1260412609 [8] WHO Monographs on Selected Medicinal Plants - Volume 1 (1999) [9] Raja AE, Vijaylakshmi M and Devalaroa G, “Acorus calamus Linn." Chemistry and Biology,” Research J. Pharm. and Tech.2. (2009) 2(2):256-261 [10] Bopana N, Saxena S, “Asparagus racemosus--Ethnopharmacological evaluation and conservation needs,” J Ethnopharmacol. (2007) 110: 1–15. Model 1 Model 2 Number of residues in favoured region (~98.0% expected) 807 (88.1%) 820 Number of residues in allowed region (~2.0% expected) 65 (7.1%) 61 (6.7%) (89.5%) Model 3 Model 4 Model 5 820 (89.5%) 54 (5.9%) 822 (89.9%) 58 (6.3%) 821 57 (6.2%) (89.8%) 18 Number of residues in outlier region Selected 44 (4.8%) 35 (3.8%) 42 (4.6%) 34 (3.7%) 36(3.9%) selected acetoxy valerenic acid bornyl_acetate β-sitosterol isovaltrate acevaltrate bornyl isovalerate calarene valeranone baldrinal 2,3-dihydro-4,5,7-trimethoxy-1- ethyl-2-methyl-3-(2,4,5-trimethoxy phenyl)indene fenchene homobaldrinal 2,4,5-trimethoxy benzaldehyde 2,6-diepishyobunone valerenal 3β, 22α, 24, 29-tetrahydroxyolean-12-en-3-O-{-β-D-arabinosyl(1→3)}-βD-arabinopyranoside valerenic acid valtroxal valepotriate valtrate xanthorrhizol 4, 5, 8-trimethoxyxanthone-2-O-β-D-glucopyranosyl(1→2)-O-β-Dgalactopyranoside Figure 1. Active constituents of Valeriana wallichii acoradin 1α, 2β, 3γ, 19α-tetrahydroxyurs-12en-28-oicacid-28-O{-β-Dglucopyranosyl (1→2)} β- D- galactopyranoside acoragermacrone Acoramone(1,2,4 –trimethoxy-5(2-propanoyl) benzene 19 β-sitosterol Calamusenone asparagamine racemosol cis-asarone(cis-1,2,4 –trimethoxy-5(2- propenyl)benzene sarsasapogenin Galangin isoeugenol methyl ether γ-cis-asarone(cis-1,2,4 –trimethoxy-5(2propenyl)benzene Isocalamendiol Limonene shatavarin Figure 3. Active constituents of Asparagus racemosus Preisocalamendiol Shyobunone Thujane Z-3-(2,4,5-trimethoxy phenyl)-2-propenal Figure 4. Ramachandran plot of COMT protein model #2 or 5. Figure 2. Active constituents of Acorus calamus 20 Figure 5. COMT protein model #2 or 5 (visualization in SPDBV) Figure 9. Interaction of COMT protein with calamusenone from Acorus calamus Figure 6. Ramachandran Plot of Grm3 mutation protein model #4 Figure 10. Interaction of COMT protein with racemosol from Asparagus racemosus Figure 7. GRM3 protein model #4(visualization if SPDBV) Figure 11. Interaction of GRM3 protein with homobaldrine from Valeriana wallichii Figure 8. Interaction of COMT protein with baldrinal from Valeriana wallichii 21 Figure 12. Interaction of GRM3 protein with galangin from Acorus calamus Figure 13. Interaction of GRM3 protein with racemosol from Asparagus racemosus 22