Akhir I nisiasi. (a) Temperatur 310K dan (b)Temperatur 312K
KESI M PULAN DAN SARAN
5.1 Kesimpulan
Berdasarkan hasil molecular docking, uji toksisitas dan simulasi dinamika molekul dapat disimpulkan bahwa CILRC merupakan kandidat antiviral terbaik untuk menghambat neuraminidase virus H5N1. Hasil simulasi molecular docking menunjukkan bahwa ligan tersebut memiliki nilai ǻ*binding sebesar -40,3614 kkal/mol dan berdasarkan hasil uji toksisitas ligan tersebut tidak memiliki potensi mutagenik, karsinogenik dan iritan.
Hasil evaluasi simulasi dinamika molekul menunjukkan bahwa ligan CILRC memiliki konformasi yang stabil pada temperatur 310K dan 312K. Ligan CILRC pada temperatur 310K membentuk ikatan hidrogen dengan Glu257, Glu258, Glu258, Arg273, Lys412, sedangkan pada temperatur 312K ikatan hidrogen yang terbentuk antara lain : Glu99, Glu99, Arg132, Trp159, Glu208, Tyr324, Arg348, Arg348, Tyr382, Glu405, Glu405. Perbedaan interaksi selama simulasi menunjukkan pengaruh temperatur terhadap aktifitas enzim.
5.2 Saran
Perlu dilakuan studi lebih lanjut secara secara wet-experiment, baik in vitro maupun in vivo. Selain untuk melihat interaksi ligan terhadap enzim
neuraminidase virus H5N1 tetapi juga untuk mengetahui dampak atau pengaruh senyawa tersebut bagi kesehatan penderita, serta untuk melihat adanya
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Universitas Indonesia Lampiran 1. Bagan Alir Penelitian
Identifikasi enzim target
Pengumpulan data sekuens enzim neuraminidase H5N1
Perancangan ligan peptida siklis disulfida Pensejajaran sekuens
Homology modeling
Struktur 3D enzim neuraminidase H5N1
Optimisasi struktur Optimisasi struktur
Target situs pengikatan Library struktur ligan
Simulasi molecular docking
Analisis hasil docking penentuan kandidat ligan terbaik Kompleks enzim neuraminidase
H5N1dengan ligan Optimisasi kompleks
Uji toksisitas Simulasi dinamika molekul
Analisis hasil dinamika molekul Kandidat antiviral H5N1
Universitas Indonesia Lampiran 2. Data Sekuen Neuraminidase ADB07955.1
neuraminidase [I nfluenza A virus (A/chicken/West Java/1074/2003 (H5N1))] GenBank: ADB07955.1
FASTAGraphics
Go to:
LOCUS ADB07955 441 aa linear VRL 01-MAR-2012
DEFINITION neuraminidase, partial [Influenza A virus (A/chicken/West
Java/1074/2003 (H5N1))]. ACCESSION ADB07955
VERSION ADB07955.1 GI:283051054 DBSOURCE accession GU183467.1 KEYWORDS .
SOURCE Influenza A virus (A/chicken/West Java/1074/2003 (H5N1))
ORGANISM Influenza A virus (A/chicken/West Java/1074/2003
(H5N1))
Viruses;; ssRNA negative-strand viruses;; Orthomyxoviridae;;
Influenzavirus A. REFERENCE 1 (residues 1 to 441)
AUTHORS Dharmayanti,N.L.P.I., Indriani,R., Darminto,D., Ibrahim,F. and
Soebandrio,A.
TITLE Genetic character of H5N1 avian influenza (AI) virus isolated from
AI vaccination flock in Indonesia JOURNAL Unpublished
REFERENCE 2 (residues 1 to 441)
AUTHORS Dharmayanti,N.L.P.I., Indriani,R., Darminto,D., Ibrahim,F. and
Soebandrio,A. TITLE Direct Submission
JOURNAL Submitted (06-NOV-2009) Department Virology, Indonesian Research
Center Institute For Veterinary Science/Bbalitvet, Jl. R.E.
Martadinata 30, Bogor, West Java 16114, Indonesia COMMENT Method: conceptual translation.
FEATURES Location/Qualifiers source 1..441
/organism="Influenza A virus (A/chicken/West Java/1074/2003 (H5N1))" /strain="A/chicken/West Java/1074/2003" /serotype="H5N1" /host="chicken" /db_xref="taxon:691001" /segment="6" /country="Indonesia" /collection_date="Nov-2003"
Universitas Indonesia Protein 1..>441 /product="neuraminidase" Region 82..414 /region_name="Sialidase"
/note="Sialidases or neuraminidases function to bind and
hydrolyze terminal sialic acid residues from various
glycoconjugates as well as playing roles in pathogenesis,
bacterial nutrition and cellular interactions. They have a
six-bladed, beta-propeller fold...;; cd00260" /db_xref="CDD:29333"
Site order(98,131,258,273,348,382,405) /site_type="active"
/note="catalytic site [active]" /db_xref="CDD:29333" Site order(238..240,242..244) /site_type="other" /note="Asp-box motif" /db_xref="CDD:29333" CDS 1..441 /gene="NA" /coded_by="GU183467.1:1..>1322" ORIGIN
1 mnpnqkiiti gsicmvigiv slmlqignmi siwvshsiqt gnqhqaesis ntnpltekav
61 asvtlagnss lcpirgwavh skdnsirigs kgdvfvirep fiscshlecr tffltqgall
121 ndkhsngtvk drsphrtlms cpvgeapspy nsrfesvaws asachdgtsw ltigisgpdn
181 gavavlkyng iitdtikswr nnilrtqese cacvngscft vmtdgpsngq asykifkmek
241 gkvvksveld apnyhyeecs cypdageitc vcrdnwhgsn rpwvsfnqnl eyqigyicsg
301 vfgdnprpnd gtgscgpmsp ngaygvkgfs fkygngvwig rtkstnsrsg femiwdpngw
361 tgtdssfsvk qdivaitdws gysgsfvqhp eltgldcirp cfwvelirgr pkestiwtsg
421 ssisfcgvns dtvswswpdg a //
Universitas Indonesia Lampiran 3. Hasil BLAST Sekuen Neuraminidase ADB07955.1
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Universitas Indonesia Lampiran 4. Multiple Sequemce Alignment dengan ClustalW2
Universitas Indonesia Lampiran 5. Ramachandran Plot
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Evaluation of residues
Residue [ 99 :GLU] ( 63.15, 44.46) in Allowed region Residue [ 105 :SER] (-104.86,-158.87) in Allowed region Residue [ 147 :PRO] ( -65.04, 106.17) in Allowed region Residue [ 202 :ASN] (-146.22, 76.45) in Allowed region Residue [ 203 :ILE] ( 63.43, 68.89) in Allowed region Residue [ 206 :THR] (-122.02,-149.56) in Allowed region Residue [ 228 :ASN] (-145.31, 20.64) in Allowed region Residue [ 276 :TRP] ( -94.50, -65.07) in Allowed region Residue [ 277 :HIS] (-155.59, 34.59) in Allowed region Residue [ 287 :ASN] ( -65.63,-177.02) in Allowed region Residue [ 324 :TYR] ( 71.62,-161.90) in Allowed region Residue [ 380 :SER] (-116.46,-138.19) in Allowed region Residue [ 435 :TRP] (-163.57,-168.39) in Allowed region Residue [ 181 :GLY] (-175.08, 38.04) in Outlier region
Number of residues in favoured region(~98.0% expected):363( 96.3%) Number of residues in allowed region (~2.0% expected) : 13( 3.4%) Number of residues in outlier region : 1( 0.3%)
Universitas Indonesia Lampiran 6. Kombinasi Ligan Pentapetida Siklis
Polar - Polar - Polar
YYY YTY YSY YKY YHY YQY YEY YCY YDY YNY YRY YYT YTT YST YKT YHT YQT YET YCT YDT YNT YRT YYS YTS YSS YKS YHS YQS YES YCS YDS YNS YRS YYK YTK YSK YKK YHK YQK YEK YCK YDK YNK YRK YYH YTH YSH YKH YHH YQH YEH YCH YDH YNH YRH YYQ YTQ YSQ YKQ YHQ YQQ YEQ YCQ YDQ YNQ YRQ YYE YTE YSE YKE YHE YQE YEE YCE YDE YNE YRE YYC YTC YSC YKC YHC YQC YEC YCC YDC YNC YRC YYD YTD YSD YKD YHD YQD YED YCD YDD YND YRD YYN YTN YSN YKN YHN YQN YEN YCN YDN YNN YRN YYR YTR YSR YKR YHR YQR YER YCR YDR YNR YRR TYY TTY TSY TKY THY TQY TEY TCY TDY TNY TRY TYT TTT TST TKT THT TQT TET TCT TDT TNT TRT TYS TTS TSS TKS THS TQS TES TCS TDS TNS TRS TYK TTK TSK TKK THK TQK TEK TCK TDK TNK TRK TYH TTH TSH TKH THH TQH TEH TCH TDH TNH TRH TYQ TTQ TSQ TKQ THQ TQQ TEQ TCQ TDQ TNQ TRQ TYE TTE TSE TKE THE TQE TEE TCE TDE TNE TRE TYC TTC TSC TKC THC TQC TEC TCC TDC TNC TRC TYD TTD TSD TKD THD TQD TED TCD TDD TND TRD TYN TTN TSN TKN THN TQN TEN TCN TDN TNN TRN TYR TTR TSR TKR THR TQR TER TCR TDR TNR TRR SYY STY SSY SKY SHY SQY SEY SCY SDY SNY SRY SYT STT SST SKT SHT SQT SET SCT SDT SNT SRT SYS STS SSS SKS SHS SQS SES SCS SDS SNS SRS SYK STK SSK SKK SHK SQK SEK SCK SDK SNK SRK SYH STH SSH SKH SHH SQH SEH SCH SDH SNH SRH SYQ STQ SSQ SKQ SHQ SQQ SEQ SCQ SDQ SNQ SRQ SYE STE SSE SKE SHE SQE SEE SCE SDE SNE SRE SYC STC SSC SKC SHC SQC SEC SCC SDC SNC SRC SYD STD SSD SKD SHD SQD SED SCD SDD SND SRD SYN STN SSN SKN SHN SQN SEN SCN SDN SNN SRN SYR STR SSR SKR SHR SQR SER SCR SDR SNR SRR KYY KTY KSY KKY KHY KQY KEY KCY KDY KNY KRY KYT KTT KST KKT KHT KQT KET KCT KDT KNT KRT KYS KTS KSS KKS KHS KQS KES KCS KDS KNS KRS KYK KTK KSK KKK KHK KQK KEK KCK KDK KNK KRK KYH KTH KSH KKH KHH KQH KEH KCH KDH KNH KRH KYQ KTQ KSQ KKQ KHQ KQQ KEQ KCQ KDQ KNQ KRQ KYE KTE KSE KKE KHE KQE KEE KCE KDE KNE KRE
Universitas Indonesia (lanjutan) KYC KTC KSC KKC KHC KQC KEC KCC KDC KNC KRC KYD KTD KSD KKD KHD KQD KED KCD KDD KND KRD KYN KTN KSN KKN KHN KQN KEN KCN KDN KNN KRN KYR KTR KSR KKR KHR KQR KER KCR KDR KNR KRR HYY HTY HSY HKY HHY HQY HEY HCY HDY HNY HRY HYT HTT HST HKT HHT HQT HET HCT HDT HNT HRT HYS HTS HSS HKS HHS HQS HES HCS HDS HNS HRS HYK HTK HSK HKK HHK HQK HEK HCK HDK HNK HRK HYH HTH HSH HKH HHH HQH HEH HCH HDH HNH HRH HYQ HTQ HSQ HKQ HHQ HQQ HEQ HCQ HDQ HNQ HRQ HYE HTE HSE HKE HHE HQE HEE HCE HDE HNE HRE HYC HTC HSC HKC HHC HQC HEC HCC HDC HNC HRC HYD HTD HSD HKD HHD HQD HED HCD HDD HND HRD HYN HTN HSN HKN HHN HQN HEN HCN HDN HNN HRN HYR HTR HSR HKR HHR HQR HER HCR HDR HNR HRR QYY QTY QSY QKY QHY QQY QEY QCY QDY QNY QRY QYT QTT QST QKT QHT QQT QET QCT QDT QNT QRT QYS QTS QSS QKS QHS QQS QES QCS QDS QNS QRS QYK QTK QSK QKK QHK QQK QEK QCK QDK QNK QRK QYH QTH QSH QKH QHH QQH QEH QCH QDH QNH QRH QYQ QTQ QSQ QKQ QHQ QQQ QEQ QCQ QDQ QNQ QRQ QYE QTE QSE QKE QHE QQE QEE QCE QDE QNE QRE QYC QTC QSC QKC QHC QQC QEC QCC QDC QNC QRC QYD QTD QSD QKD QHD QQD QED QCD QDD QND QRD QYN QTN QSN QKN QHN QQN QEN QCN QDN QNN QRN QYR QTR QSR QKR QHR QQR QER QCR QDR QNR QRR EYY ETY ESY EKY EHY EQY EEY ECY EDY ENY ERY EYT ETT EST EKT EHT EQT EET ECT EDT ENT ERT EYS ETS ESS EKS EHS EQS EES ECS EDS ENS ERS EYK ETK ESK EKK EHK EQK EEK ECK EDK ENK ERK EYH ETH ESH EKH EHH EQH EEH ECH EDH ENH ERH EYQ ETQ ESQ EKQ EHQ EQQ EEQ ECQ EDQ ENQ ERQ EYE ETE ESE EKE EHE EQE EEE ECE EDE ENE ERE EYC ETC ESC EKC EHC EQC EEC ECC EDC ENC ERC EYD ETD ESD EKD EHD EQD EED ECD EDD END ERD EYN ETN ESN EKN EHN EQN EEN ECN EDN ENN ERN EYR ETR ESR EKR EHR EQR EER ECR EDR ENR ERR CYY CTY CSY CKY CHY CQY CEY CCY CDY CNY CRY CYT CTT CST CKT CHT CQT CET CCT CDT CNT CRT CYS CTS CSS CKS CHS CQS CES CCS CDS CNS CRS
Universitas Indonesia (lanjutan)
CYK CTK CSK CKK CHK CQK CEK CCK CDK CNK CRK CYH CTH CSH CKH CHH CQH CEH CCH CDH CNH CRH CYQ CTQ CSQ CKQ CHQ CQQ CEQ CCQ CDQ CNQ CRQ CYE CTE CSE CKE CHE CQE CEE CCE CDE CNE CRE CYC CTC CSC CKC CHC CQC CEC CCC CDC CNC CRC CYD CTD CSD CKD CHD CQD CED CCD CDD CND CRD CYN CTN CSN CKN CHN CQN CEN CCN CDN CNN CRN CYR CTR CSR QKR CHR CQR CER CCR CDR CNR CRR DYY DTY DSY DKY DHY DQY DEY DCY DDY DNY DRY DYT DTT DST DKT DHT DQT DET DCT DDT DNT DRT DYS DTS DSS DKS DHS DQS DES DCS DDS DNS DRS DYK DTK DSK DKK DHK DQK DEK DCK DDK DNK DRK DYH DTH DSH DKH DHH DQH DEH DCH DDH DNH DRH DYQ DTQ DSQ DKQ DHQ DQQ DEQ DCQ DDQ DNQ DRQ DYE DTE DSE DKE DHE DQE DEE DCE DDE DNE DRE DYC DTC DSC DKC DHC DQC DEC DCC DDC DNC DRC DYD DTD DSD DKD DHD DQD DED DCD DDD DND DRD DYN DTN DSN DKN DHN DQN DEN DCN DDN DNN DRN DYR DTR DSR DKR DHR DQR DER DCR DDR DNR DRR NYY NTY NSY NKY NHY NQY NEY NCY NDY NNY NRY NYT NTT NST NKT NHT NQT NET NCT NDT NNT NRT NYS NTS NSS NKS NHS NQS NES NCS NDS NNS NRS NYK NTK NSK NKK NHK NQK NEK NCK NDK NNK NRK NYH NTH NSH NKH NHH NQH NEH NCH NDH NNH NRH NYQ NTQ NSQ NKQ NHQ NQQ NEQ NCQ NDQ NNQ NRQ NYE NTE NSE NKE NHE NQE NEE NCE NDE NNE NRE NYC NTC NSC NKC NHC NQC NEC NCC NDC NNC NRC NYD NTD NSD NKD NHD NQD NED NCD NDD NND NRD NYN NTN NSN NKN NHN NQN NEN NCN NDN NNN NRN NYR NTR NSR NKR NHR NQR NER NCR NDR NNR NRR RYY RTY RSY RKY RHY RQY REY RCY RDY RNY RRY RYT RTT RST RKT RHT RQT RET RCT RDT RNT RRT RYS RTS RSS RKS RHS RQS RES RCS RDS RNS RRS RYK RTK RSK RKK RHK RQK REK RCK RDK RNK RRK