Table S1. Distribution of GAD and ADC homologs in Archaea Phylum Order Species GAD type ADC type Aeropyrum camini YP_008603414.1 Aeropyrum pernix NP_146906.2 Desulfurococcus fermentans YP_006401609.1 Desulfurococcus kamchatkensis YP_002427717.1 Desulfurococcus mucosus YP_004175993.1 Hyperthermus butylicus YP_001012441.1 Desulfurococcales Ignicoccus hospitalis Ignisphaera sedula Pyrolobus fumarii Staphylothermus hellenicus YP_003669158.1 Staphylothermus marinus YP_001041291.1 Thermogladius cellulolyticus YP_006362587.1 Thermosphaera aggregans YP_003649423.1 Acidianus hospitalis Metallosphaera cuprina Metallosphaera sedula Crenarchaeota Sulfolobales Metallosphaera yellowstonensis Sulfolobus acidocaldarius YP_255707.1 Sulfolobus islandicus Sulfolobus solfataricus Sulfolobus tokodaii Caldivirga maquilingensis Pyrobaculum aerophilum Pyrobaculum arsenaticum YP_001153708.1 Pyrobaculum calidifontis Pyrobaculum islandicum YP_930884.1 Pyrobaculum neutrophilum YP_001793753.1 Thermoproteales Pyrobaculum oguniense YP_005260639.1 YP_005260632.1 Thermofilum pendens Thermoproteus neutrophilus Thermoproteus tenax YP_004892846.1 Thermoproteus uzoniensis Vulcanisaeta distributa Vulcanisaeta moutnovskia YP_004244042.1 Archaeoglobus fulgidus NP_070828.1 Archaeoglobales Archaeoglobus profundus YP_003400310.1 Archaeoglobus veneficus YP_004341026.1 Ferroglobus placidus YP_003436047.1 Haladaptatus paucihalophilus WP_018128932.1 Halalkalicoccus jeotgali YP_003737798.1 Haloarchaeum acidiphilum WP_021779700.1 Haloarcula amylolytica WP_008312494.1 Haloarcula argentinensis WP_005537837.1 Haloarcula hispanica YP_004796901.1 Haloarcula japonica WP_004591335.1 Haloarcula marismortui YP_136395.1 Haloarcula sinaiiensis WP_004957873.1 Haloarcula vallismortis WP_004517137.1 Halobacterium salinarum NP_279424.1 Halobiforma lacisalsi WP_007143761.1 Halobiforma nitratireducens WP_006672080.1 Halococcus morrhuae WP_004051068.1 Halococcus saccharolyticus WP_006076597.1 Halococcus salifodinae WP_005039049.1 Halococcus thailandensis WP_007739300.1 Haloferax alexandrinus WP_006600569.1 Haloferax denitrificans WP_004971205.1 Haloferax elongans WP_008323373.1 Haloferax gibbonsii WP_004972627.1 Haloferax larsenii WP_007545249.1
Euryarchaeota Halobacteriales Methanobacteriales Methanocellales Haloferax mediterranei Haloferax mucosum Haloferax prahovense Haloferax sulfurifontis Haloferax volcanii Halogeometricum borinquense Halogranum salarium Halomicrobium katesii Halomicrobium mukohataei Halopiger xanaduensis Haloquadratum walsbyi Halorhabdus tiamatea Halorhabdus utahensis Halorubrum aidingense Halorubrum californiense Halorubrum coriense Halorubrum hochstenium Halorubrum kocurii Halorubrum lacusprofundi Halorubrum lipolyticum Halorubrum saccharovorum Halorubrum tebenquichense Halosarcina pallida Haloterrigena limicola Haloterrigena thermotolerans Haloterrigena salina Haloterrigena turkmenica Halosimplex carlsbadense Halovivax asiaticus Halovivax ruber Natrialba aegyptia Natrialba asiatica Natrialba chahannaoensis Natrialba hulunbeirensis Natrialba magadii Natrialba taiwanensis Natrinema altunense Natrinema gari Natrinema pallidum Natrinema pellirubrum Natrinema versiforme Natronobacterium gregoryi Natronococcus amylolyticus Natronococcus jeotgali Natronococcus occultus Natronomonas moolapensis Natronomonas pharaonis Natronolimnobius innermongolicus Natronorubrum bangense Natronorubrum sulfidifaciens Natronorubrum tibetense Methanobacterium formicicum Methanobrevibacter ruminantium Methanobrevibacter smithii Methanosphaera stadtmanae Methanothermobacter marburgensis Methanothermobacter thermautotrophicus Methanothermus fervidus Methanocella arvoryzae Methanocella conradii Methanocella paludicola Methanocaldococcus fervens Methanocaldococcus infernus Methanocaldococcus jannaschii YP_006348490.1 WP_008321575.1 WP_008092283.1 WP_007275119.1 YP_003534871.1 YP_004037409.1 WP_009366427.1 WP_018257853.1 YP_003178807.1 YP_004598387.1 YP_008377911.1 YP_003131637.1 WP_007998188.1 WP_008440485.1 WP_006114288.1 WP_008581765.1 WP_008849150.1 YP_002565263.1 WP_008004675.1 WP_004049554.1 WP_006629814.1 WP_008389243.1 WP_008008967.1 WP_006650054.1 WP_008896200.1 YP_003402374.1 WP_006885814.1 WP_007697606.1 YP_007283870.1 WP_006664550.1 WP_006110079.1 WP_006169253.1 WP_006654055.1 YP_003480720.1 WP_006826692.1 WP_007107678.1 WP_008457034.1 WP_006186465.1 YP_007282015.1 WP_006429090.1 YP_007178631.1 WP_005553684.1 WP_008427146.1 YP_007308260.1 YP_007488063.1 YP_326257.1 WP_007258363.1 WP_006066558.1 WP_008162697.1 WP_006089115.1 WP_004030562.1 YP_003424638.1 WP_019264882.1 YP_447382.1 YP_003850392.1 WP_010876740.1 YP_004003689.1 YP_686121.1 YP_005379982.1 YP_003355454.1 YP_003128590.1 YP_003615689.1 NP_247014.1 YP_658098.1
Korarchaeota Thaumarchaeota Nanoarchaeota Methanocaldococcus sp. FS406-22 YP_003457883.1 Methanocaldococcus villosus WP_004589931.1 Methanocaldococcus vulcanius YP_003247431.1 Methanococcales Methanococcus aeolicus YP_001325198.1 Methanococcus maripaludis NP_987251.1 Methanococcus vannielii YP_001323648.1 Methanococcus voltae YP_003707961.1 Methanothermococcus thermolithophicus WP_018153725.1 Methanothermococcus okinawensis YP_004576297.1 Methanotorris igneus YP_004484887.1 Methanotorris formicicus WP_007043501.1 Methanocorpusculum labreanum YP_001030976.1 Methenoculleus bourgensis YP_006544180.1 Methanoculleus marisnigri YP_001047756.1 Methanofolis liminatans WP_004037849.1 Methanolinea tarda WP_007313863.1 Methanomicrobiales Methanoplanus limicola WP_004076822.1 Methanoplanus petrolearius YP_003893553.1 Methanoregula boonei YP_001405323.1 Methanoregula formicica YP_007248156.1 Methanosphaerula palustris YP_002467101.1 Methanospirillum hungatei YP_504027.1 Methanopyrales Methanopyrus kandleri NP_614783.1 Methanococcoides burtonii YP_566374.1 Methanohalobium evestigatum YP_003727876.1 Methanohalophilus mahii YP_003543170.1 Methanomethylovorans hollandica YP_007314131.1 Methanosaeta concilii YP_004385069.1 Methanosarcinales Methanosaeta harundinacea YP_005919754.1 Methanosaeta thermophila YP_844043.1 Methanosalsum zhilinae YP_004617076.1 Methanosarcina acetivorans NP_614980.1 Methanosarcina barkeri YP_304529.1 Methanosarcina mazei NP_633341.1 Pyrococcus abyssi NP_126894.1 Pyrococcus furiosus YP_006492243.1 Pyrococcus horikoshii NP_142859.1 Pyrococcus sp. NA2 YP_004424537.1 Pyrococcus sp.st04 YP_006354801.1 Pyrococcus yayanosii YP_004623572.1 Thermococcus barophilus YP_004072136.1 Thermococcales Thermococcus gammatolerans YP_002960503.1 Thermococcus kodakarensis YP_184227.1 Thermococcus litoralis YP_008428447.1 Thermococcus onnurineus YP_002308097.1 Thermococcus sibiricus YP_002993875.1 Thermococcus zilligii WP_010479297.1 Thermococcus sp. 4557 YP_004762928.1 Thermococcus sp. AM4 YP_002582098.1 Thermococcus sp. CL1 YP_006424975.1 Ferroplasma acidarmanus YP_008142735.1 Thermoplasmatales Picrophilus torridus YP_022928.1 Thermoplasma acidophilum Thermoplasma volcanium Candidatus Korarchaeum cryptofilum Cenarchaeales Cenarchaeum symbiosum Nitrosopumilales Nitrosopumilus maritimus Nanoarchaeum equitans Green colored boxes indicate the presence of a homolog on the genome, along with the accession number. White colored boxes indicate their absence.
Aful ------------------------------------------------------------ Saci ------------------------------------------------------------ Ptor ------------------------------------------------------------ Mmaz ------------------------------------------------------------ Mbur ------------------------------------------------------------ Mthp ------------------------------------------------------------ Mhun ------------------------------------------------------------ Hsal ------------------------------------------------------------ Mkan ------------------------------------------------------------ Mjan ------------------------------------------------------------ Mmar ------------------------------------------------------------ Mtht ------------------------------------------------------------ Paby ------------------------------------------------------------ Phor ------------------------------------------------------------ Pfur ------------------------------------------------------------ Tkod ------------------------------------------------------------ Hs67 MASSTPSSSATSSNAEPDPNTTNLRPTTYDTWCGVAHGCTRKLGLKICGFLQRTNSLEEK 60 Mm67 MASSTP-SPATSSNAGADPNTTNLRPTTYDTWCGVAHGCTRKLGLKICGFLQRTNSLEEK 59 Hs65 MASPGS---GFWSFGSEDGSGDSENPGTARAWCQVAQKFTGGIGNKLCALLYGD---AEK 54 Mm65 MASPGS---GFWSFGSEDGSADPENPGTARAWCQVAQKFTGGIGNKLCALLYGD---SGK 54 Aful ------------------------------------------------------------ Saci ------------------------------------------------------------ Ptor ------------------------------------------------------------ Mmaz ------------------------------------------------------------ Mbur ------------------------------------------------------------ Mthp ------------------------------------------------------------ Mhun ------------------------------------------------------------ Hsal ------------------------------------------------------------ Mkan ------------------------------------------------------------ Mjan ------------------------------------------------------------ Mmar ------------------------------------------------------------ Mtht ------------------------------------------------------------ Paby ------------------------------------------------------------ Phor ------------------------------------------------------------ Pfur ------------------------------------------------------------ Tkod ------------------------------------------------------------ Hs67 SRLVSAFKERQSSKNLLSCENSDRDARFRRTETDFSNLFARDLLPAKNGEEQTVQFLLEV 120 Mm67 SRLVSAFRERQSSKNLLSCENSDQGARFRRTETDFSNLFAQDLLPAKNGEEQTAQFLLEV 119 Hs65 PAESGGSQPPRAAARKAACACDQKPCSCSKVDVNYAFLHATDLLPACDGERPTLAFLQDV 114 Mm65 PAEGGGSVTSRAATGKVACTCDQKPCNCPKGDVNYAFLHATDLLPACDGERPTLAFLQDV 114
Aful --MDSSSVDIVLNKGGHNKSFEVKIYRIHTTKIMSFPNGSDAEGVLKRLEDYAKNDFEPH 58 Saci ---MKAFPDKPLSKQ--------DIYEIAKT--------------------YSRNDNEPL 29 Ptor --MLKRFPERGIPLD--------EINSILDG--------------------YGKNDIKNS 30 Mmaz ------MNEQGLSEREIF---------------------------------SYLENAKSE 21 Mbur ------MEENGKTKEEIL---------------------------------LFLKKAKSA 21 Mthp --MMYTFPEKGLSEDMVT---------------------------------DLLKEMRSR 25 Mhun ------MDAEGLSTDELF---------------------------------CFLQAKRNE 21 Hsal -----------MTRG----------------------------------------EARRP 9 Mkan -------------------------------------------------------MILQR 5 Mjan ---MRNMQEKGVSEKEIL---------------------------------EELKKYRSL 24 Mmar -----------MDEQDIL---------------------------------NELREYRNQ 16 Mtht ------------------------------------------------------------ Paby ---MSKFPEKGLPREEVL---------------------------------NLLEDKTKV 24 Phor ----MKFPRIGLPKEKVI---------------------------------ELINEKTKK 23 Pfur ----MKFPRKGIPQEEVM---------------------------------RELEKYTSK 23 Tkod -----MFPERGASEEEVL---------------------------------RELEEKTRE 22 Hs67 VDILLNYVRKTFDRSTKVLDFHHPHQLLEGMEGFNLELSDHPESLEQILVDCRDTLKYGV 180 Mm67 VDILLNYVRKTFDRSTKVLDFHHPHQLLEGMEGFNLELSDHPESLEQILVDCRDTLKYGV 179 Hs65 MNILLQYVVKSFDRSTKVIDFHYPNELLQ---EYNWELADQPQNLEEILMHCQTTLKYAI 171 Mm65 MNILLQYVVKSFDRSTKVIDFHYPNELLQ---EYNWELADQPQNLEEILTHCQTTLKYAI 171 Aful SRRMWGHIYYAGL-KDVVELARKAYLMYMDKTMLDFTCFPSLLRMEREVVRMASSLLNGD 117 Saci SGRMWGHIYSLGLPEDVIEVSMTLYNQFINKTMLDFTVYPSVLRFENDIIAMASSLLGGN 89 Ptor RGRLFTYFYDPGL-KDLDDLS-SILLKFYNRNGMDYHAFPSTLKIENDLISMMSDLMHGN 88 Mmaz DTDYYR--VFSSMCTRPHKIAIEANRLFIEANLGDLGLFAGAHKLEQEVVRMLGNLLHAS 79 Mbur DASYER--VLSSMCTYPHEIAVLAHTQFIESNMGDPGLFPGTFNLEKQVLAMFGKMLHHK 79 Mthp DCPYDR--LLSTMCTRPHPVAVRAYSMFLETNLGDPGLFPGTAEIERRVVGILGSLLGCS 83 Mhun DFSYSH--ILSSMCTTPHPVAVQAHNLFMETNLGDPGLFPGTATLEDRLIRWFADLYHEP 79 Hsal PQEFDR--VLSSMCTTPHPAAREAAQAFLATNPGDPETYPAVAERERDAVALLGEIVGLS 67 Mkan DSDYSDGTVLGSMCTEPHPVAAEAFVAGLHVNLGDPYLFPNAYRAERECIGWLAETLLDH 65 Mjan DLKYEDGNIFGSMCSNVLPITRKIVDIFLETNLGDPGLFKGTKLLEEKAVALLGSLLN-- 82 Mmar DLKYEEGYILGSMCTKPHPMARKISEMFFETNLGDPGLFKGTSKLEKEVVSMIGGILH-- 74 Mtht -MTYTSGRILGSMCTSSHPLARRVYCDFLESNLGDPGLFRGTRELESGVIGMLGELLS-- 57 Paby DLTFSSGKILGSMCTMPHELAIEVFARYIDRNLGDPGLHPGTRKIEEEVIEMLSDLLH-- 82 Phor DLTFSSGKILGSMCTMPHDLAIEVYTKYIDRNLGDPGLHPGTRKIEEEVIEMISDLLH-- 81 Pfur DLSFSSGKILGSMCTLPHELAKEVFCMYMDRNLGDPGLHPGTKKIEEEVIEMLSDLLH-- 81 Tkod DLTFDSGKILGSMCTYPHPFAVKVVMKYIDRNLGDPGLHIGSQKIEKEAVDMLANLLG-- 80 * * * Hs67 RTGHPRFFNQLSTGLDIIGLAGEWLNSTANTNMFTYEIAPVFVLMEQITLKKMREIVGWS 240 Mm67 RTGHPRFFNQLSTGLDIIGLAGEWLTSTANTNMFTYEIAPVFVLMEQITLKKMREIVGWS 239 Hs65 KTGHPRYFNQLSTGLDMVGLAADWLTSTANTNMFTYEIAPVFVLLEYVTLKKMREIIGWP 231 Mm65 KTGHPRYFNQLSTGLDMVGLAADWLTSTANTNMFTYEIAPVFVLLEYVTLKKMREIIGWP 231
Aful ---EEVVG---------NFTYGGTESIMLALKAAREKFRKEEGG-----NVVPEIVLPAT 160 Saci ---EETVG---------NFTFGGTESIMVATKSARDYFLKRHS------SVIPEILLPVT 131 Ptor ---DDTSG---------TFTTGGTESILLAMKAARDLFLEKKE-------YVPEIVAPVT 129 Mmaz S-IDVPSGGLCQSSVCGYLTTGGTESNIQAVRGMKNLVTAGKKE----FKGTPNIVIPAS 134 Mbur N-SPEKAG---------YLTTGGTESNIQAIRSMHNFRHDISR---------PNIVMPES 120 Mthp ----DATG---------YVSTGGTESNIQAVRAARNSSGRRDG----------NIVVPRS 120 Mhun ----SAGG---------CTTSGGTESNIQVLRFCKKTKNVKEP----------NIIVPAS 116 Hsal ----SPHG---------YIAAGGTEANLQAVRAARNRADADAV----------NVVAPAS 104 Mkan PAPEEAEG---------SIVSGGTEANILAAYAAREVTG----------GR--EIIVPAT 104 Mjan --NKDAYG---------HIVSGGTEANLMALRCIKNIWREKRRKG-LSKNEHPKIIVPIT 130 Mmar --NKNAFG---------YLISGGTEANLTAMRAFKNISK--------SKGKPQNIIIPET 115 Mtht --EPDAAG---------HIITGGTEANLMAMRAARNMAG----------AEKPEIIVPKS 96 Paby --LEKGYG---------HIVSGGTEANILAVRAFRNISD----------AERPELILPKS 121 Phor --LEKGHG---------HIVSGGTEANILAVRAFRNLSD----------VEKPELILPKS 120 Pfur --LERGYG---------HIVSGGTEANILAVRAFRNLAD----------VENPELILPKS 120 Tkod --LEKGYG---------HIVSGGTEANILAVRAMRNLAG----------IEKPELILPES 119 * **** * * ** Hs67 S--KDGDG---------IFSPGGAISNMYSIMAARYKYFPEVKTKGMAAVPKLVLFTSEQ 289 Mm67 N--KDGDG---------IFSPGGAISNMYSIMAARYKYFPEVKTKGMAAVPKLVLFTSEH 288 Hs65 G--GSGDG---------IFSPGGAISNMYAMMIARFKMFPEVKEKGMAALPRLIAFTSEH 280 Mm65 G--GSGDG---------IFSPGGAISNMYAMLIARYKMFPEVKEKGMAAVPRLIAFTSEH 280 Aful AHPAFWKSAEYLGMRCLRAKLDDELR------ADVETVKELVGDK---TAMIVGSAPNYP 211 Saci AHPAFNKASDYLGMKVTPVKIDPERTT-----VDLEDLKSKLKEN---TAMIVASAPNYP 183 Ptor AHPAFSKAAKYLGMKITRVPVNEDYI-------ADDTINEYINDR---TAAVIASAPSFP 179 Mmaz AHFSFDKVADMMGIEVRRASLDSEFR------VDMASVEKLINEN---TIGLVGIAGNTE 185 Mbur AHFSFDKVANLSGIEIRKASLDKLLK------VDLDSVRSLIDKN---TIGLVGIAGTTE 171 Mthp AHFSFDKIADLLNLEVRKAELDESLR------VDVGDVERLIDDR---TVCLVGIAGTTE 171 Mhun AHFSFEKACGMMDIEMRVVPVDEQYR------MKTDAAGELIDKN---TCCIVGVAGTTE 167 Hsal AHFSFQKAADVLGVELRLAPTDGDHR------ADVAAVADLVDGD---TAVVVGVAGTTE 155 Mkan RHFSFEKAARMLRMKLVEAPLRSDYT------VDVDAVQDLISRD---TALIVGIVGTTE 155 Mjan AHFSFEKGREMMDLEYIYAPIKEDYT------IDEKFVKDAVED--YDVDGIIGIAGTTE 182 Mmar AHFSFDKAKDMMDLNVVRPPLTKYFT------MDVKFIKDYIEDSKNEVSGIVGIAGCTE 169 Mtht AHFSFRKAADILGLRLREAELDQDYR------VDVESVRKLISEN---TVAVVGVAGTTE 147 Paby AHFSFIKAGEMLGVKLVWAELKQDYA------VDVKDVEAKISDN---TIGIVGIAGTTG 172 Phor AHFSFIKAGEMLGVKLVWAELNPDYT------VDVRDVEAKISDN---TIGIVGIAGTTG 171 Pfur AHFSFIKAGEMLGVKLIWADLNPDYT------VDVKDVEAKISEN---TIGIVGIAGTTG 171 Tkod AHFSFIKAAEMLGVKLVWAELNDDYT------VNVKDVEKKITDR---TIGIVGIAGTTG 170 * * * * * Hs67 SHYSIKKAGAALGFGTDNVILIKCNERGKIIPADFEAKILEAKQKGYVPFYVNATAGTTV 349 Mm67 SHYSIKKAGAALGFGTDNVILIKCNERGKIIPADLEAKILDAKQKGYVPLYVNATAGTTV 348 Hs65 SHFSLKKGAAALGIGTDSVILIKCDERGKMIPSDLERRILEAKQKGFVPFLVSATAGTTV 340 Mm65 SHFSLKKGAAALGIGTDSVILIKCDERGKMIPSDLERRILEVKQKGFVPFLVSATAGTTV 340
Aful FGVVDDIKALSDIAVDGKLWLHVDACLGGFHLPFFRELG---EKIPDFDFSVEGVHSISA 268 Saci FGTIDDVKALSEIAQDKKLWLHVDSCIGGFLLPFLRDLG---EPIPPFDLSLEGVTSISA 240 Ptor YGGIDNIKDISEIALDKNTWFHVDACVGGMILPFLKGLG---LNIKDFDFKLPGVSSMSI 236 Mmaz FGQIDPIDKLSEVALENELFLHVDAAFGGFVIPFLEKPQ-------PFDFKVPGVTSIAI 238 Mbur FGQLDPINELSKIAIEKGIFLHIDAAFGGFVIPFMDIDY-------TYDFRLEGVTSMTI 224 Mthp FGQVDPIGDLSELAIENGIPLHVDAAFGGFVLPFLEKDC-------MWDFRAEGVQSITI 224 Mhun YGMTDPIPALGKLAEQEGVHLHVDAAFGGYVLPFLDDAP-------PFDFSVPGVGSIAV 220 Hsal YGRVDPIPALADIAAGVDANLHVDAAWGGFVLPFTDHD---------WSFADAPVNTMAI 206 Mkan TGSVDDIEALSDVAEDHGVPLHVDAAFGGFTAPFLREE----YPLPRFGFDLEAVVSVTV 211 Mjan LGTIDNIEELSKIAKENNIYIHVDAAFGGLVIPFLDDKYKKKGVNYKFDFSL-GVDSITI 241 Mmar LGSIDNICELSKIAVENDILLHVDAAFGGFVIPFLDDKYKLDGYNYDFDFSLNGVSSITI 229 Mtht LGRIDPVEELSEICLDEDIHLHIDAAFGGFIIPFLRETG---AELPEFDFKLQGVSSITV 204 Paby LGVVDDIPALSDLAREYGIPLHVDAAFGGFVIPFAKSLG---YDLPDFDFKLKGVESITI 229 Phor LGVVDDIPALSDLARDYGIPLHVDAAFGGFVIPFAKELG---YELPDFDFKLKGVQSITI 228 Pfur LGVVDDIPALSDLARDYGIPLHVDAAFGGFVIPFAKELG---YDLPDFDFKLKGVQSITI 228 Tkod LGVVDDIPALSDLALDYGLPLHVDAAFGGFVIPFAKALG---YEIPDFDFRLKGVKSITI 227 * * * * ** ** * * * * * ** Hs67 YGAFDPIQEIADICEKYNLWLHVDAAWGGGLLMSRKHRHK--------LNGIERANSVTW 401 Mm67 YGAFDPIQEIADICEKYNLWLHVDAAWGGGLLMSRKHRHK--------LSGIERANSVTW 400 Hs65 YGAFDPLLAVADICKKYKIWMHVDAAWGGGLLMSRKHKWK--------LSGVERANSVTW 392 Mm65 YGAFDPLLAVADICKKYKIWMHVDAAWGGGLLMSRKHKWK--------LSGVERANSVTW 392 Aful DFHKYGLSPRGASVILYRNAKLREGQIFVMASWPGYPLVN-TAVLSTRSAGTLAA----- 322 Saci DLHKYGYAPRGASVVLFRNSSYREGSIFVMSRWPGYPIVN-TSVLSTRSAGPLAA----- 294 Ptor DLHKYGFTPKGSSVVLYKNHDLRKRQIYVNADWPGYPMSN-MGMQATKSAGPLAG----- 290 Mmaz DPHKMGLSTIPSGALLFRSPSFLDSLKVSTPYLT--TKSQ-FTLTGTRSGASAAA----- 290 Mbur DPHKMALSTIPSGGLLFKEPEYFECLEIHTPYLS--VNKQ-YSLTGTRSGAGVAS----- 276 Mthp DPHKMGMSPIPAGGLIFRSSDPLRRLETETYYLT--VSRQ-ASLTGTRSGAAAAA----- 276 Mhun DPHKMGLSTIPSGVLMVRDERVFCNLLVETPYLT--TKQA-YSLTGTRPGASVAA----- 272 Hsal DPHKMGQAPVPAGGFLARDPETLDALAIETPYLE--SDTQ-PTLGGTRSGAGVAG----- 258 Mkan DPHKMGLVPPPAGGIVFRDDEFPKAIEVYAPYLSGGGASQ-YTITGTRPGAPVLA----- 265 Mjan DPHKMGHCPIPSGGILFKDIGYKRYLDVDAPYLT--ETRQ-ATILGTRVGFGGAC----- 293 Mmar DPHKMGLAPISAGGILFRDNMFKKYLDVDAPYLT--EKQQ-ATIIGTRSGVGVAS----- 281 Mtht DPHKMGLAPIPSGCILFRDASYLDAMSIETPYLT--EKQQ-STIVGTRTGASAAA----- 256 Paby DPHKMGMAPIPAGGIIFRRKKYLKAISVLAPYLAGGKVWQ-ATITGTRPGASVLA----- 283 Phor DPHKMGMAPIPAGGIVFRRKKYLKAISVLAPYLAGGKVWQ-ATITGTRPGASVIA----- 282 Pfur DPHKMGMAPIPAGGIVFRHKKYLRAISVLAPYLAGGKIWQ-ATITGTRPGASVLA----- 282 Tkod DPHKMGMVPIPAGGIIFREKKFLDSISVLAPYLAGGKIWQ-ATITGTRPGANALA----- 281 * ** * ** * Hs67 NPHKMMGVLLQCSAILVKEKGILQGCNQMCAGYLFQPDKQYDVSYDTGDKAIQCGRHVDI 461 Mm67 NPHKMMGVLLQCSAILVKEKGILQGCNQMCAGYLFQPDKQYDVSYDTGDKAIQCGRHVDI 460 Hs65 NPHKMMGVPLQCSALLVREEGLMQNCNQMHASYLFQQDKHYDLSYDTGDKALQCGRHVDV 452 Mm65 NPHKMMGVPLQCSALLVREEGLMQSCNQMHASYLFQQDKHYDLSYDTGDKALQCGRHVDV 452
Aful --AWAVMSYLGFDGYLKLAKKTLYAKKRLIDGLTELG-LELLG--SPEGAVLAFTSERH- 376 Saci --AWGIIHGLGKDGYRKLANRILNTRIKLTNELPKMG-YRILG--KPLGGIVSFTSEEF- 348 Ptor --SWATLNYLGLDGYKKLAEKTLKAYRMIRSGITDLG-YKIIG--RPDATIFAFTHNDK- 344 Mmaz --TCAVMKYLGYEGYRKNVQYCMELTSKIVEEARKLG-FEPLI--EPVMNVVALKVPN-- 343 Mbur --TYAVMKHLGRKGYKKVVSDCMSVTKKLVDGAEKLG-INTVI--DPVLNIVALDVPE-- 329 Mthp --TYAVIMHLGIDGYRKVVRRCMDMTEHLVSEARAMG-IEPVI--EPVMNVVALRVDD-- 329 Mhun --AYAVMAYLGRKGMKALVTGCMENTRRMIEGMEAFG-VHRKV--TPDVNVATFEHVS-- 325 Hsal --ALASLRALWPDGYREQYERTQGNAEYLAAELAARG-YD-VV--DPELPLVAADMPD-- 310 Mkan --LYANILELGEEGYRRIAFRCYEETLKVAEKARELGLELAVD--PPHLNLVNIRLPDRG 321 Mjan --TYAVLRYLGREGQRKIVNECMENTLYLYKKLKENNFKPVI---EPILNIVAIEDEDY- 347 Mmar --TWGIMKLLGIDGYETLVNESMEKTMYLVKKAREYGFETAI---DPVMNIVALNDENK- 335 Mtht --TWAIMKHMGREGYRKLALRVMGVTRRLRDGLVELDYQLVV---EPELNIVAFNHPAMG 311 Paby --VWALIKHLGFEGYREIVRKAMELSRWFAEEIKKLNNAWLVR--EPMLNIVSFQTKN-- 337 Phor --VWALIKHLGFEGYMRIVERAMKLSRWFAEEIKKINNAWLVR--EPMLNIVSFQTKN-- 336 Pfur --VWALIKHLGFEGYMEIVDRAMKLSRWFAEEIKKTPGAWLVR--EPMLNIVSFKTKN-- 336 Tkod --VWAMIKHLGFDGYKEVVKEKMELARWFASELKKIPGIYLIR--EPVLNIVSFGSEK-- 335 * * * * Hs67 FKFWLMWKAKGTVGFENQINKCLELAEYLYAKIKNREEFEMVFNGEPEHTNVCFWYIPQ- 520 Mm67 FKFWLMWKAKGTVGFENQINKCLELADYLYAKIKNREEFEMVFDGEPEHTNVCFWYIPQ- 519 Hs65 FKLWLMWRAKGTTGFEAHVDKCLELAEYLYNIIKNREGYEMVFDGKPQHTNVCFWYIPP- 511 Mm65 FKLWLMWRAKGTTGFEAHIDKCLELAEYLYTIIKNREGYEMVFDGKPQHTNVCFWFVPP- 511 Aful NLFKVSTLMAEKGWYVQSQPGSKKLGFPRSLHFSVIPGHAEVVDEFLEDMREVLPECECS 436 Saci NLAELPTLM--KGWFIQYQPGSRILGFPKSIHLTIAPGHDKVVDEFLRDLERATIELRGK 406 Ptor DIIDLGIKMIENGWYPQIQPGNVFIDMPDSVHLNISPVHLDVADEFLEFFNELDKNVKPR 404 Mmaz PDLVRERLLKKFGWNVSITRTPRALRLVLMPHNSPED-----IELFLEDLKKVTAEIKSP 398 Mbur ADLVRKKLLDEYGWHVSITRNPRALRIVIMPHIKNET-----IELFLKDLAKVIK----- 379 Mthp PPGVRRALLER-GWHVSMTREPKALRLILMPHMTDEN-----LDLFLSDLEDVLISLRRG 383 Mhun VPSP---------WVVSYTRK-GDLRIVCMPHVTRDV-----VEAFLSDFGESYVSHIS- 369 Hsal AEFQALR---EEGWRISRTAS-DALRVVCMPHVTREM-----LAAFLDDVDALA------ 355 Mkan TAERLLRESEREGWKISVSTKPLGVRIVMMPHLDAET-----VSRFLELVARVLGG---- 372 Mjan --KEVCKKLRDRGIYVSVCNCVKALRIVVMPHIKREH-----IDNFIEILNSIKRD---- 396 Mmar --HDTCMKLRDENWYVSVCRCVDALRIVVMPHLEIEH-----IDGFLESLSNTKKY---- 384 Mtht -PHELADRLEELGWAVSVSSCPPAIRVVLMPHIMEEH-----IELLLRDLEGIRLREE-- 363 Paby -LRKVERELKRRGWGISAHRG--YIRIVFMPHVTKEM-----VEEFLRDLREVLK----- 384 Phor -LKKVERELKSRGWGISAHRG--YIRIVFMPHVTREM-----IEEFLKDLKEVLS----- 383 Pfur -LRRVERELKSRGWGISAHRG--YIRIVS--HASCDG-----GHD--------------- 371 Tkod -LEELEKELKARGWGVSAHRG--YIRIVVMPHVKREH-----LEEFLRDLREIAKRL--- 384 * Hs67 SLRGVPDSPQRREKLHKVAPKIKALMMESGTTMVGYQPQGDKANLFRMVISNPAATQSDI 580 Mm67 SLRGVPDSPERREKLHRVAPKIKALMMESGTTMVGYQPQGDKANFFRMVISNPAATQSDI 579 Hs65 SLRTLEDNEERMSRLSKVAPVIKARMMEYGTTMVSYQPLGDKVNFFRMVISNPAATHQDI 571 Mm65 SLRTLEDNEERMSRLSKVAPVIKARMMEYGTTMVSYQPLGDKVNFFRMVISNPAATHQDI 571
Aful ---YPEMSSFD-----VSKLKFGEDGLPEDSELISELIHSMPPEIVESVFKQFINELIFR 488 Saci RLTLPPLQDFNDLQSIARALGIEEGKLPSNPTLINELMHEMPPELVENVLKLVINEYVFR 466 Ptor -----DKNSYN-----VNSVEKALEMIKMEKSMLFRIIRYSRPEVSEKIFLEMTDEDFTY 454 Mmaz ------------------------------------------------------------ Mbur ------------------------------------------------------------ Mthp G----------------------------------------------------------- 384 Mhun ------------------------------------------------------------ Hsal ------------------------------------------------------------ Mkan ------------------------------------------------------------ Mjan ------------------------------------------------------------ Mmar ------------------------------------------------------------ Mtht ------------------------------------------------------------ Paby ------------------------------------------------------------ Phor ------------------------------------------------------------ Pfur ------------------------------------------------------------ Tkod ------------------------------------------------------------ Hs67 DFLIEEIERLGQDL---------------------------------------------- 594 Mm67 DFLIEEIERLGQDL---------------------------------------------- 593 Hs65 DFLIEEIERLGQDL---------------------------------------------- 585 Mm65 DFLIEEIERLGQDL---------------------------------------------- 585 Aful ---- Saci PSRS 470 Ptor S--- 455 Mmaz ---- Mbur ---- Mthp ---- Mhun ---- Hsal ---- Mkan ---- Mjan ---- Mmar ---- Mtht ---- Paby ---- Phor ---- Pfur ---- Tkod ---- Hs67 ---- Mm67 ---- Hs65 ---- Mm65 ----
Figure S1. Sequence alignment of GAD type proteins. The primary structures of 16 representative archaeal GAD homologs (upper sequences) and 4 mammalian GAD proteins (lower sequences) were aligned with the ClustalW program provided by the DNA Data Bank of Japan (DDBJ). Amino acid residues that are conserved only in archaeal GAD homologs are indicated with asterisks and in red, and those shared by archaeal GAD homologs and mammalian GADs are indicated with asterisks and in blue. Residue numbers are indicated on the right. Abbreviations represent the proteins from the following organisms: Aful (Archaeoglobus fulgidus), Saci (Sulfolobus acidocaldarius), Ptor (Picrophilus torridus), Mmaz (Methanosarcina mazei), Mbur (Methanococcoides burtonii), Mthp (Methanosaeta thermophila), Mhun (Methanospirillum hungatei), Hsal (Halobacterium salinarum), Mkan (Methanopyrus kandleri), Mjan (Methanocaldococcus jannaschii), Mmar (Methanococcus maripaludis), Mtht (Methanothermobacter thermautotrophicus), Paby (Pyrococcus abyssi), Phor (Pyrococcus horikoshii), Pfur (Pyrococcus furiosus), Tkod (Thermococcus kodakarensis), Hs67 (human GAD67), Mm67 (mouse GAD67), Hs65 (human GAD65) and Mm65 (mouse GAD65).
Figure S2 Tomita et al M 6 mer 2 mer (kda) 97 66 45 30 20 Legend for reviewers Fig. S2. SDS-PAGE analysis of the purified, recombinant TK1814 protein. Three micrograms of hexameric and dimeric TK1814 proteins were applied and the gels were stained with Coomassie Brilliant Blue.
Figure S3 Tomita et al Specific activity (nmol min -1 mg -1 ) 700 600 500 400 300 200 100 0 5 6 7 8 9 10 11 ph Fig. S3. The effects of ph on the aspartate decarboxylase activity of the TK1814 protein. Symbols: open circles, MES (ph 5.5-7.0); open squares, PIPES (ph 6.5-7.5); triangles, HEPES (ph 7.0-8.0); diamonds, Bicine (ph 8.0-9.0); closed circles, Tricine (ph 8.0-9.0); closed squares, CHES (ph 9.0-10.0).
Figure S4 Tomita et al. (A) PpyrF pyrf pud3/ 1814 KUW1 1814 single crossover insertion PpyrF 1814 pyrf 1814 pyrf PpyrF pop-out recombination 1814 (B) outside ( ) inside ( ) M host Δ host Δ Legend for reviewers Fig. S4. Disruption of the TK1814 gene. (A) The plasmid and recombination strategy to construct the TK1814 gene disruption strain. T. kodakarensis KUW1 (Δ pyrf, Δ trpe) was used as the host strain. Cells that had undergone an initial single crossover insertion were enriched by growing cells in medium without uracil. Cells that had undergone a second, pop-out recombination were selected with resistance toward 5-FOA. (B) PCR analyses of the TK1814 locus. The positions of the primers used are indicated with arrowheads in (A).