Supplemental Figure Legend

Fig. S1. Control gel electrophoretic analysis of free Ph-P0 samples. The samples (each

including 100 pmol of Ph-P0 or its mutants) of Ph-P0.Ph-L12 complex (lane 1), WT Ph-P0

alone (lane 2), Ph-C∆79 alone (lane 3), Ph-C∆89 alone (lane 4), Ph-C∆105 alone (lane 5),

Ph-C∆114 alone (lane 6), Ph-C∆131 alone (lane 7), together with 800 pmol of Ph-L12 alone

(lane 8), were analyzed by the same gel as in Fig. 2A.

Fig. S2. SDS-PAGE analysis of the reconstituted complexes appeared as shifted bands in

Fig. 2A. The shifted bands in Fig. 2A (lane 2 to 6) were cut out and applied on the

SDS-PAGE (lane 4 to 8, respectively). The band of free Ph-L12 not shifted (lane 7, Fig.2A)

was also cut out and analyzed in lane 9. WT Ph-P0 alone (lane 2, 100pmol), and Ph-L12 alone

(lane 3, 600pmol) were applied on the same gel. The MW ladder marker was loaded on lane 1.

The characteristic smearing patterns of Ph-L12, as shown in lanes 3 and 9, appeared in all the

lanes of complexes (lane 4 to 8). The Ph-L12 area of smearing was indicated in the right side

of gel panel. The band of WT Ph-P0 (closed circle) or its mutants (closed triangle) was also

detected in individual lanes. These results confirm the complex formation between Ph-L12

and Ph-P0 variants except C∆131.

Fig. S3. Binding of Ph-P0.Ph-L12 complexes to the 50S core particles deficient in

L10.L7/L12 and L11. The 50S core (10 pmol, lane 1) were incubated with Ph-P0.Ph-L12

complexes (20 pmol each), which were reconstituted using WT-P0 (lane 2), C∆79 (lane 3),

C∆89 (lane 4), C∆105 (lane 5), C∆114 (lane 6), and C∆131 (lane 7). The samples were

analyzed by electrophoresis on an acrylamide-agarose composite gel. After electrophoresis the

gel was stained with Azur B.

Fig. S4. Complex formation of [32P]-labeled Ph-L12 with Ph-P0 or its truncation mutants.

(A) Ph-L12 was [32P]-phosphorylated in vitro by casein kinase II as described previously (31).

The labeled Ph-L12 (800 pmol; 42 cpm/pmol of protein) was used in the formation of

Ph-P0.Ph-L12 complex with 100 pmol of WT-P0 (lane 1), C∆79 (lane 2), C∆105 (lane 3) or

C∆131 (lane 4). After native gel electrophoresis of the samples, the gel was stained with

Coomassie Brilliant Blue. (B) The same gel as in A was subjected to autoradiography. As

shown in A and B, a significant amount of 32P-labeled Ph-L12 formed the complex with WT

Ph-P0 (lane 1). However, reduced amounts of the labeled proteins formed the complex with

C∆79 (lane 2) and an even lower amount with C∆105 (lane 3). Only free 32P-Ph-L12 was

detected with the C∆131 sample (lane 4). The ratio of radioactivity of the complexes

containing WT Ph-P0, C∆79 and C∆105 was 2.6: 1.9: 1.

Fig. S5. Alignment of the amino acid sequences of P0 and L10. The amino acid sequences of

P0 from 11 species of Eukarya and 10 species of Archaea, and of L10 from 28 species of

Bacteria were aligned by clustalW algorithm followed by manual inspection. The alignment is

separated by spaces into three domains, upper, middle and lower, corresponding to the

urkingdoms, Eukarya, Archaea, and Bacteria. The positions of the N-terminal and C-terminal

regions with the symbol ‘x’ attached on the space between the archaeal and bacterial domains

could not be aligned between the eukaryal/archaeal and the bacterial sequences, while those of

the C-terminal region with the symbol ‘x’ between the eukaryal and archaeal domains could not

be aligned between the eukaryal and archaeal sequences. Selected positions used for

phylogenetic analysis were marked with ‘*’ above the alignment. The putative binding sites of

stalk dimers were masked based on the results of biochemical experiments on the silkworm

(Bmo) P0 for eukaryal and archaeal sequences (31), and on the reported structural information

for Thermotoga maritima (Tma) L10 for bacterial sequences (23). The position of the inserted

sequences is enclosed by a rectangle. In this portion all archaeal sequences have insertions

compared to eukaryotic sequences, while several bacterial sequences have insertions compared

to other bacterial sequences. The archaeal and bacterial insertions could not be aligned either.

Abbreviations: [Eukarya] Sce, Saccharomyces cerevisiae; Spo, Schizosaccharomyces pombe;

Hsa, Homo sapiens; Dma, Drosophila melanogaster; Cel, Caenorhabditis elegans; Ddi,

Dictyostelium discoideum; Ath, Arabidopsis thaliana; Tcr, Trypanosoma cruzi; Pfa,

Plasmodium falciparum; Gla, Giardia lamblia; Bmo, Bombyx mori; [Archaea] Sac, Sulfolobus

acidocaldarius; Pae, Pyrobaculum aerophilum; Ape, Aeropyrum pernix; Mva, Methanococcus

vannielii; Mka, Methanopyrus kandleri; Afu, Archaeoglobus fulgidus; Hma, Haloarcula

marismortui; Tac, Thermoplasma acidophilum; Pto, Picrophilus torridus; Pho, Pyrococcus

horikoshii; [Bacteria] Tma, Thermotoga maritima; Eco, Escherichia coli; Sau, Staphylococcus

aureus; Bsu, Bacillus subtilis; Smu, Streptococcus mutans; Ljo, Lactobacillus johnsonii; Mpu,

Mycoplasma pulmonis; Aae, Aquifex aeolicus; Tth, Thermus thermophilus; Dra, Deinococcus

radiodurans; Cte, Clostridium tetani; Fnu, Fusobacterium nucleatum; Mle, Mycobacterium

leprae; Cdi, Corynebacterium diphtheriae; San, Streptomyces antibioticus; Nsp, Nostoc sp; Gvi,

Gloeobacter violaceus; Lin, Leptospira interrogans; Rpr, Rickettsia prowazekii; Atu,

Agrobacterium tumefaciens; Hpy, Helicobacter pylori; Cje, Campylobacter jejuni; Tpa,

Treponema pallidum; Bbu, Borrelia burgdorferi; Neu, Nitrosomonas europaea; Nme, Neisseria

meningitides; Paer, Pseudomonas aeruginosa; Bba, Bdellovibrio bacteriovorus.

1 2 3 4 5 6 7 8

Free Ph-P0

Supplemental Fig. S1

Supplemental Fig. S2

1 2 3 4 5 6 7 8 9

30kDa

20kDa

15kDa

35kDa

10kDa

Ph-P0 WT

Ph-P0 C∆ mutants

Ph-L12

Supplemental Fig. S3

Supplemental Fig. S4

Fig. S5

1.......10........20........30........40........50........60........70........80........90.......100.......110.......120 ** *** *********************************** *************** ** ** Sce ------------------MGGIREKKA-EYF-AKLREYLEEYKSLFVVGVDNVSSQQMHEVRKELRGR----AVVLMGKNTMVRRAIRG-FLS---DL--P--DFEKLLP-------FVK 81 Spo ------------------MAISKESKA-QYF-EKLRSLFEKYNSLFVVNIDNVSSQQMHTVRKQLRGT----AELIMGKNTMIRRAMRG-IIN---DM--P--ELERLLP-------VVR 81 Hsa ----------------MPREDRATWKS-NYF-LKIIQLLDDYPKCFIVGADNVGSKQMQQIRMSLRGK----AVVLMGKNTMMRKAIRG-HLE---NN--P--ALEKLLP-------HIR 83 Dme ----------------MVRENKAAWKA-QYF-IKVVELFDEFPKCFIVGADNVGSKQMQNIRTSLRGL----AVVLMGKNTMMRKAIRG-HLE---NN--P--QLEKLLP-------HIK 83 Cel ----------------MVREDRSTWKA-NYF-TKLVELFEEYPKCLLVGVDNVGSKQMQEIRQAMRGH----AEILMGKNTMIRKALRG-HLG---KN--P--SLEKLLP-------HIV 83 Ddi -----------------MSGAGSKRKK-LFI-EKATKLFTTYDKMIVAEADFVGSSQLQKIRKSIRGI----GAVLMGKKTMIRKVIRD-LAD---SK--P--ELDALNT-------YLK 82 Ath ---------------MAVKATKAEKKI-VYD-SKLCQLLNEYSQILVVAADNVGSTQLQNIRKGLRGD----SVVLMGKNTMMKRSVRI-HAD---KTGNQ--AFLSLLP-------LLQ 86 Tcr ------------------MPSVSEAKR-EYE-ERFNGCLTKYGRVLFCLMDNVRSQQVHDVRRDLRGL----GELVMGKKTLQKKIVER-RAE---DKKAS--AYDKLLYNTCIEKKLLC 90 Pfa ----------------MAKLSKQQKKQ-MYI-EKLSSLIQQYSKILIVHVDNVGSNQMASVRKSLRGK----ATILMGKNTRIRTALKK-NLQ---AV--P--QIEKLLP-------LVK 83 Gla -------------MSAAPLTAKQARRQ-AYV-AKLERCLTEYKKIVLVSVDNVRSFQIAQIRRLLRGK----AELLAGKNTIIKRVINQ-LDD-------D--KLKNLLP-------YIK 84 Bmo ----------------MGREDKATWKS-NYF-VKIIQLLDEYPKCFIVGADNVGSQQMQQIRISLRGS----SIVLMGKNTMMRKAIKD-HLD---NN--P--ALEKLLP-------HIK 83 Sac ----------MIGLAVTTTKKIAKWKV-DEV-AELTEKLKTHKTIIIANIEGFPADKLHEIRKKLRGK----ADIKVTKNNLFNIALKN-AG--------Y--DTKLFES-------YLT 86 Pae ------MMLAIGKRRYVRTRQYPARKV-KIV-SEATELLQKYPYVFLFDLHGLSSRILHEYRYRLRRY----GVIKIIKPTLFKIAFTK-VYG-------G--IPAEIAE-------KVR 91 Ape -----MSVVSLVGQMYKREKPIPEWKT-LML-RELEELFSKHRVVLFADLTGTPTFVVQRVRKKLWKK----YPMMVAKKRIILRAMKA-AGL---E---L--DDNLLDD-------LVR 93 Mva ------------MIDAKSEHKIAPWKI-EEV-NALKELLKSANVIALIDMMEVPAVQLQEIRDKIRDQ----MTLKMSRNTLIKRAVEE-VAE---ETGNP--EFAKLVD-------YLD 89 Mka -----MAVKAKGQPPSGYEPKVAEWKR-REV-KELKELMDEYENVGLVDLEGIPAPQLQEIRAKLRER---DTIIRMSRNTLMRIALEE-KLD---ER--P--ELEPLLD-------YIE 95 Afu --------------MAAVRGSPPEYKV-RAV-EEIKRMISSKPVVAIVSFRNVPAGQMQKIRREFRGK----AEIKVVKNTLLERALDA-LG--------G--DYLKLKD-------YLG 82 Hma ----------MSAESERKTETIPEWKQ-EEV-DAIVEMIESYESVGVVNIAGIPSRQLQDMRRDLHGT----AELRVSRNTLLERALDD-VD--------D--GLEDLNG-------YIT 86 Tac ------------------MKEVSQQKK-ELV-NEITQRIKASRSVAIVDTAGIRTRQIQDIRGKNRGK----INLKVIKKTLLFKALEN-LG-------DE--KLSKLKE-------NSG 79 Pto ------------------MTEPAQWKI-DFV-KNLENEINSRKVAAIVSIKGLRNNEFQKIRNSIRDK----ARIKVSRARLLRLAIEN-TG-------KN--NIVKLKD-------YAH 79 Pho ------------------MAHVAEWKK-KEV-EELAKLIKSYPVIALVDVSSMPAYPLSQMRRLIREN---GGLLRVSRNTLIELAIKK-AAK---ELGKP--ELEKLVE-------YID 84 xxxxxxxxxxxxxxxxxxxxxxxxx Tma -------------------MLTRQQKE-LIV-KEMSEIFKKTSLILFADFLGFTVADLTELRSRLREKYGDGARFRVVKNTLLNLALKN-AEY---EGYEE--FLKGPTA-------VLY 86 Eco ------------------MALNLQDKQ-AIV-AEVSEVAKGALSAVVADSRGVTVDKMTELRKAGREA---GVYMRVVRNTLLRRAVEG-TPF---ECLKD--AFVGPTL-------IAY 84 Sau ------------------MSAIIEAKK-QLV-DEIAEVLSNSVSTVIVDYRGLTVAEVTDLRSQLREA---GVEYKVYKNTMVRRAAEK-AGI---EGLDE--FLTGPTA-------IAT 84 Bsu ------------------MSSAIETKK-VVV-EEIASKLKESKSTIIVDYRGLNVSEVTELRKQLREA---NVESKVYKNTMTRRAVEQ-AEL---NGLND--FLTGPNA-------IAF 84 Smu -----------------MSEASIAKKA-ELV-DAVAEKMKAAVSIVVVDSRGLTVEQDTVLRRNLRES---AVEFKVIKNSILRRAAEK-AGL---EGFDD--IFTGPSA-------VAF 85 Ljo -----------------MSKAIIAKKE-KLV-DDFAAELKEAKAILVIDYLGLTVEEVTNLRKDLRDA---NVKMKVIKNTYLKRAAEK-AGI---EGLED--TFVGPTA-------VVY 85 Mpu ---------------MHIESKNRQIKK-DIV-VEITKKINDSQSLFLAEYRGLTVAKLLNLRKEAKKH---NVEIKVYKNRLVKLATQK-LGH---SDLDK--FLVGPNL-------FVF 87 Aae MAEFDK-EAYAERERSAASRKTLLKKQ-ELVNSYKERLQKSNGFVIFFNFQGIDAYPLTLLRLDIKDL---KGEIVVGKNTLFYRAFSD-TVL---SDHRD--IFVGPTA-------ALF 102 Tth -------------------MPNKRNVE-LLA-TLKENLERAQGSFFLVNYQGLPAKETHALRQALKQN---GARLFVAKNTLIRLALKE-LGL----PELD--GLQGPSA-------VVF 82 Dra -------------------MANEKNQQ-TLG-SLKDSLQGIE-TFYVVDYQGLTAGQLTQLRKDIREK---GGQLIVAKNTLLNLALQE-GGR----DFDD--ALKGPSA-------LVL 81 Cte ------------MRRWHTVSKNRQLKE-AKV-AEIKEKMEKAECIVLADYQGLTVEEATELRKKMREE---GVEYKVFKNTLSILAAKE-LGY---EGITD--LFQGPIS-------IAF 90 Fnu --------------------MATQVKK-ELV-AELVEKIKKAQSVVFVDYQGIKVNEETLLRKQMREN---GAEYLVAKNRLFKIALKE-SGV--EDSFDE--ILEGSTA-------FAF 83 Mle --------------------MARADKA-TAV-ANIAEQFKAATAALITEYRGLTVANLAELRRSLTG----SASYAVAKNTLIKRAASE-VGI---EGLDE--LFAGPTA-------IAF 81 Cdi --------------------MANPKNT-ESL-AELKNRFADIDSVFVTEYRGLTVAQITELRRALGS----DVQYSVAKNTLIKLAAKE-AGI---EGLDD--ILTGPTA-------VAF 81 San --------------------MARPDKA-AAV-AELTDKFRSSNAAVLTEYRGLTVAQLKTLRRSLGE----NAQYAVVKNTLTKIAAKE-AGI---TLEDQ--LFAGPTA-------VAF 81 Nsp ------------------MGRTLENKK-EIV-ADLKETLGESTLALVIEYQGLTVAEITDLRKRLRPS---GTVCKVTKNTLMGIAIQDDEKW---QPLSE--LLKGSSA-------FLL 85 Gvi ------------------MGKR-PVKE-ALV-GDLQKLLEKSSVVMVIDYRGLSVAEITGLRRRMREH---GGTCVVAKNTLMGVATRE-TAW---RNIDP--LLAGPSA-------FLF 83 Lin --------------------MANQEKI-EAV-ALLKGKLETRNNFILACYSGLTVEEITGLRAQLRKE---GSEMKVLKNNLFLRALKE-SGAHKDKNITFGSEYQGPLA-------AIF 87 Rpr --------------------MLRSEKP-VAV-EDIVNIYKESPSVIITHYHGLTVSQVSSLREELKSK---EAGFKVVKNTLAKIAAKQ-TGL---DSITN--LFAGPTA-------IVY 82 Atu --------------------MERAEKR-EFV-TELNEVFKASGSVVVAHYAGVTVAQMNDFRSKMRAA---GGTVKVAKNRLAKIALQG-TES---EGMTN--LFKGQTL-------IAY 82 Hpy ----------------MQKQHQRQHKV-ELV-ANLKSQFADAKALLICDYKGLSVRKLEALRNKARNQ---GIKVQVIKNTLAHIAMKE-TGY----SDLD---LKETNV-------FLW 84 Cje --------------------MTRSEKV-EII-AKLEEGFKASEAIVVCNYRGLSTKKLEELRNNAREN---NVKVQIVKNTLANIALNN-SGK----TGLV---LKDTNI-------YLW 80 Tpa --------------MAVRARRLQPAKV-AAV-ESLTRDLGEASSYIFTEYRGLTVEQLTALRRALREF---SCVYRVVRNNFANIAFTS-LNM----TVGE--YLVGPTA-------IAL 87 Bbu ------------------MSAKINAKKLEMF-DLLKQFIDSKQNLFFLDYRGLNVAQLTELRNKIEGE---HGSLKVVKNNIMKMVLKE-KNI---NVVDS--CLVGPTV-------VVT 85 Neu ------------------MSHNLEGKK-AIV-AEVSSQVAKAQAIVIAEYRGLGVDQFTRLRVKARES---GIYFRVIKNTFARRAVAD-TPF---SGLAE--SMVGPLA-------YGI 84 Nme ------------------MSLNIETKK-VAV-EEISAAIANAQTLVVAEYRGISVSSMTELRANARKE---GVYLRVLKNTLARRAVQG-TSF---AELAD--QMVGPLV-------YAA 84 Paer ------------------MAIKLEDKK-AIV-AEVNEAAKAALSAVVADARGVTVGAMTGLRKEAREA---GVYVKVVRNTLLKRAVEG-TQF---DVLND--VFKGPTL-------IAF 84 Bba -------------MKGGSHMITRADKE-QEI-KLITEKFGKAKGAFIVDFKGIKVEQVTNLRKKLNAA---DSEMKVVRNTLAKRAFKDHPAI--ETAFAN--SMKGTNA-------IVF 91

.......130.......140.......150.......160.......170.......180.......190.......200.......210.......220.......230.......240 * **** ******** **************** Sce GYV-GFVFTNEPLTEIKNVIVSNRVAA--PARAGAVAPEDIWVRAVNTGMEPG-KTSFFQALGVPTKIARGTIEIVSDVKVVDAGNKVGQSEASLLNLLNISPFTFGLTVVQVYDNGQVF 197 Spo GNV-GFVFTNADLKEVRETIIANVIAA--PARPNAIAPLDVFVPAGNTGMEPG-KTSFFQALGIPTKITRGTIEITSDVHLVSKDAKVGPSEATLLNMLNISPFTYGMDVLTIYDQGNVF 197 Hsa GNV-GFVFTKEDLTEIRDMLLANKVPA--AARAGAIAPCEVTVPAQNTGLGPE-KTSFFQALGITTKISRGTIEILSDVQLIKTGDKVGASEATLLNMLNISPFSFGLVIQQVFDNGSIY 199 Dme GNV-GFVFTKGDLAEVRDKLLESKVRA--PARPGAIAPLHVIIPAQNTGLGPE-KTSFFQALSIPTKISKGTIEIINDVPILKPGDKVGASEATLLNMLNISPFSYGLIVNQVYDSGSIF 199 Cel ENV-GFVFTKEDLGEIRSKLLENRKGA--PAKAGAIAPCDVKLPPQNTGMGPE-KTSFFQALQIPTKIARGTIEILNDVHLIKEGDKVGASESALLNMLGVTPFSYGLVVRQVYDDGTLY 199 Ddi QNT-CIIFCKDNIAEVKRVINTQRVGA--PAKAGVFAPNDVIIPAGPTGMEPT-QTSFLQDLKIATKINRGQIDIVNEVHIIKTGQKVGASEATLLQKLNIKPFTYGLEPKIIYDAGACY 198 Ath GNV-GLIFTKGDLKEVSEEVAKYKVGA--PARVGLVAPIDVVVQPGNTGLDPS-QTSFFQVLNIPTKINKGTVEIITPVELIKKGDKVGSSEAALLAKLGIRPFSYGLVVESVYDNGSVF 202 Tcr GNT-ALIFTNEEIPVITAVLDKHRVQA--PARVGP-SPMRRHCPAGNTGMEPK-ATSFFQALNIATKIAKGTVEIVSDKKVLSVGDRVDNSTATLLQKLDISPFYYQVEVQSVWDRGMLF 205 Pfa LNM-GFVFCKDDLSEIRNIILDNKSSSH-PARLGVIAPIDVFIPPGPTGMDPS-HTSFLESLGISTKIVKGQIEIQEHVHLIKQGEKVTASSATLLRKFNMNP-SYGVDVRTVYDDGVIY 199 Gla LNV-AFVFTNGDTSAILKAFKKTKRKA--AAKAGIVAPADVVIEPMLTQSGPD-QHGFYAALGIDTKINKGKIEIVNPVNLIKKGDIVTPSHATLLQRLEIDPFFYAMSALNLYDDGEIY 200 Bmo GNV-GFVFTRGDLVEVRDKLLENKVQA--PARPGAIAPLSVVIPAHNTGLGPE-KTSFFQALSIPTKISKGTIEIINDVHILKPGDKVGASEATLLNMLNISPFSYGLVVKQVYDSGTIF 199 Sac GPN-AFIFTDTNPFELQLFLSKFKLKR--YALPGDKADEEVVVPAGDTGIAAGPMLSVFGKLKIKTKVQDGKIHILQDTTVAKPGDEIPADIVPILQKLGIMPVYVKLNIKIAYDNGVII 203 Pae GEV-GFFFTSFNPAEVIKIVAENSVRR--AAQPGDKAPFDIVVPAGPTNASPGPIISKFGKLKIPTRVQEGKIWIAKDTVVAKAGQEITPEMAEVLRVVGIEPIFEQLRLLGVIWRGQRF 208 Ape GQM-LLVFADGNPFKIVKEVEKEKVAM--PVKPGDKAETEIRIPEGMTNLTPGPILSVFGKLRIQYQVRGGKIYIAKETVVAKPGDVISEDLAGLLMALGIRPIEKGVRVKFAIDGGVLI 210 Mva KGA-AIVVTEMNPFKLFKTLEESKSPA--PIKGGAIAPCDIEVKSGSTGMPPGPFLSELKAVGIPAAIDKGKIGIKEDKVVAKEGDVISPKLAVVLSALGIKPVTVGLNVLGVYEEGVIY 206 Mka GPV-AFIFTNLDPFKLYKLLEESKASA--PAKPGDIAPEDIVVPEGPTPFEPGPIVSELQQAGLPAQIQDGKVVITKDTVLVKEGEEIDEKTAEILKKLEIEPMEVGVDIVAIVAEGTLF 212 Afu DQI-AIITADENPFRLFRMIEDTKVPS--PLKPNQVSPVDVVVNEGPTPIPPGPLMAELQMAGLPVAIEKGKVVVKATTTVVKAGEVVRPEVARALERLDIKPIKIGLDVKAMLDSGVIL 199 Hma GQV-GLIGTDDNPFSLFQELEASKTPA--PIGAGEVAPNDIVIPEGDTGVDPGPFVGELQSVGADARIQEGSIQVLSDSTVLDTGEEVSQELSNVLNELGIEPKEVGLDLRAVFADGVLF 203 Tac GQI-ALLTSDLEPTEIYKIIESTFQKT--APRGGEIAPEDIVIQPMTTGFPPGPMMTEFQKVGLQTGVEKGKIAIKKETVFVKKGETIPKDKAKILEMLEIKPLEVGLQLLGLYSEGVVY 196 Pto GQV-ALITTDESPKKIYDILEKSKTKA--PAKGGEIAEEDIVIEPKETNFPPGPKISEFQKAGLPAAIEKGKIVIKSEVTFVKKGDVITREKALVLKDLEIYPITAGLDLIAAYEDGLIF 196 Pho RGA-GILVTNMNPFKLYKFLQQNRQPA--PAKPGAVVPKDVVVPAGPTPLAPGPIVGQMQALGIPARIEKGKVTIQKDTTVLKAGEVITPELANILNALGIQPLEVGLDVLAVYEDGIVY 201 Tma VTE-GDPVEAVK--IIYNFYKDKKAD--------------------------------------------------------------------------LSRLKGGFLEGKKFTAEEVE 129 Eco SM--EHPGAAAR--LFKEFAKANA----------------------------------------------------------------------------KFEVKAAAFEGELIPASQID 124 Sau SS--EDAVAAAK--VISGFAKDHE----------------------------------------------------------------------------ALEIKSGVMEGNVITAEEVK 124 Bsu ST--EDVVAPAK--VLNDFAKNHE----------------------------------------------------------------------------ALEIKAGVIEGKVSTVEEVK 124 Smu SN--EDVVAPAK--IINDFAKDAE----------------------------------------------------------------------------ALEIKGGAIEGAVSTKEEIQ 125 Ljo TADAEDITEPAR--IVSKYEDDID----------------------------------------------------------------------------ALSIKGGMLEGKVASQEEIK 127 Mpu SN--ELGMDGAK--VLAEFAKKNK----------------------------------------------------------------------------KLVLKAGIFDGKVVDAAGVK 127 Aae AY--EDPVAVTK--KLVEFLKETFDKEW------------------------------------------------------------------------EGRIKGGLLDYKYITPEQVK 146 Tth Y---EDPVAAAK--TLVQFAKSNPKG--------------------------------------------------------------------------IPQVKSGLLQGQILTAKDVE 123 Dra AQ--EDPAGVAK--ALSDAAGRNDRG--------------------------------------------------------------------------IPTVKGGFVEGSKVDVAVVQ 123 Cte GY--EDPTAPAR--ILNDFAKDHK----------------------------------------------------------------------------KLELKGGMVQGEVYDEDKIK 130 Fnu GY--NDPVAPAK--AVFDLAKAKAKAKLD-----------------------------------------------------------------------VFKIKGGYLTGKKVSVKEVE 128 Mle VT--GEPVDAAK--AIKTFAKEHK----------------------------------------------------------------------------ALVIKGGYMDGHPMTVAEVE 121 Cdi IK--GEAVDAAK--AMKKFASENK----------------------------------------------------------------------------AFVIKGGYMDGNALSAAQVD 121 San VT--GDPVESAK--GLRDFAKENP----------------------------------------------------------------------------NLVIKGGVLDGKALSADEIK 121 Nsp VK--EDFSSAIK--AYQEFQKVTK----------------------------------------------------------------------------KTELRGGVMEGRLLKEPDVK 125 Gvi GN--EKLKEMLK--TYEDFARETK----------------------------------------------------------------------------KTEFRGAVVDGTLVTLDGLK 123 Lin AK--DNLPAIAK--VCKDFAKNNK----------------------------------------------------------------------------NLIVKAGYMDGSVLDANGVD 127 Rpr S---KEPVEMAR--LVVNFANSND----------------------------------------------------------------------------NLKIIGGIVDNHILDAYSIK 121 Atu S---VDPMIAPK--VVMDFAKTND----------------------------------------------------------------------------KVVVLGGSMGATTLNAEAVK 121 Hpy G---GDQIALSK--LVFDFQKEHKD---------------------------------------------------------------------------HFVLKAGLFDKESVSVAHVE 124 Cje G---EDQLSVSK--VAAKFEENND----------------------------------------------------------------------------KFEIKTAHIEGEVADVAKVK 119 Tpa VDT-EHANGVAR--VLFDFAKEVP----------------------------------------------------------------------------ALVVKGAILDGEVFDASKVE 128 Bbu AL--EEANVIAK--IFYDFVKSS-----------------------------------------------------------------------------TLKVKGGFVLGEFYDEAKVQ 124 Neu G---SDPVATAK--ILHEFAKDND----------------------------------------------------------------------------RFVIKAGAMAGIVMSDKDVA 123 Nme S---EDAVAAAK--VLHQFAKKDD----------------------------------------------------------------------------KIVVKAGSYNGEVMNAAQVA 123 Paer SN--EHPGAAAR--IFREFAKGQD----------------------------------------------------------------------------KFEIKAAAFEGQFLAANQID 124 Bba SY--GEVNATAK--ALADFAKDVE----------------------------------------------------------------------------VLQIKSGVMDGAALDDAKIK 131

.......250.......260.......270.......280.......290.......300.......310.......320.......330.......340.......350.......360 *** ************ Sce -PSSILDITDEELVSHFVSAVSTIASISLAIGY-PTLPSVGHTLINNYKDLLAVAIAASY-HYPEIEDLVDRIEN-------------------PEKYAAAAPAATSAASGDAAPA---- 291 Spo -SPEILDVSEEDLIGHLLSAASIITAISLGANY-PTILSVMHSVVNAYKNLVAVSLATEY-TFEGTEQTKAFLAD-------------------PSAFVVAAAPAAAAGGEAEAPAAEAA 295 Hsa -NPEVLDITEETLHSRFLEGVRNVASVCLQIGY-PTVASVPHSIINGYKRVLALSVETDY-TFPLAEKVKAFLAD-------------------PSAFVAAAPVAAATTAAPAAAAAP-A 296 Dme -SPEILDIKPEDLRAKFQQGVANLAAVCLSVGY-PTIASAPHSIANGFKNLLAIAATTEV-EFKEATTIKEYIKD-------------------PSKFAAAASASAAPAAGGATEKKE-- 295 Cel -TPEVLDMTTEELRKRFLSGVRNVASVSLAVNY-PTLASVAHSLANGLQNMLGVAAVTDV-SFKEAETIKAFIAD-------------------PSKFAAAAPAAAAAPAAAAPAAKK-- 295 Ddi -SPS---ISEEDLINKFKQGIFNIAAISLEIGY-PTVASIPHSVMNAFKNLLAISFETSY-TFDAAEKFKSAAAA-------------------APVRAAPSAAAPRAAAKKVVV----- 288 Ath -NPEVLNLTEDDLVEKFAAGVSMITALSLAISY-PTVAAAPHMFLNAYKNVLAVALATEY-SFPQAENVKEFLKD-------------------PTKFAVAVAAPVSGESGGAVVAVAV- 299 Tcr -LREDLSITDDVVEKYLLEGISNVAALSLGAGI-PTAATLPHMIMDAFKTLLGASVATEY-EFDEFDGKNLRKAA-------------------LEGNLGGGVADAAAAADTGAAAAPAA 303 Pfa -DAKVLDITDEDILEKFSKGVSNVAALSRATGV-ITEASYPHVFVEAFKNIVALIIDSDY-TFPLMKILKKWVEN-------------------PEAFAAVAAPASAAKADEPKKEEAKK 297 Gla -DAAVLEIDDSVMEAKWNAGLEAFVSLALGANF-PCLPAIPHIFMDTAKSFIGAGVEADVTEIPLVKRVKEILAD-------------------PSKFAVAAPAAATAAAAAPEEAKK-- 297 Bmo -APEILDIKPEDLRAKFQAGVANVAALSLAIGY-PTIASAPHSIANGFKNLLAIAAVTEV-EFEEATTIKEFIKD-------------------PSKFAAVAAAVAPSAAAAPA------ 291 xxxxxxxxxxxxxxxxxxxxxxxxxx Sac -PGDKLSINLDDYTNEIRKAHINAFAVATEIAY-PEPKVLEFTATKAMRNALALASEIGY-ITQE--TAQAVFTKAVMKAYAVASSISGKVD--LGVQIQAQPQVSEQAA---------- 306 Pae VDISELIIDVNKYKELFETASVYARNLALNIVY-PTREVLQAVIPAAHMRAVALAAKLGV-VTRE--TLPALLSRAVAEANALAAVVAAKAPD-LGISVSLPQTAAPQQTPQPTEAP--- 320 Ape -TEDLLRPDIEAFRGDVIDAAKEALGLATEIVYMPVPEAVESAIVKAALAASALAAETGF-IAPG--TVEDVVRKAIAEEAAVVALLGDKARE-LGIEEAAPAAAPAAEEKAEEEKK--- 322 Mva -TSDVLRIDEEEFLGKLQKAYTNAFNLSVNAVI-PTSATIETIVQKAFNDAKAVSVESAF-ITEK--TADAILGKAHAQMIAVAKLAGDEA---LDDDLKEQISSSAVVATEEAPKAET- 317 Mka -ERDDLAIDFDEYEDMAKEAAQHAFNLSINAAI-PTAETADVIVAKAHTEALNLAVNAGV-PVPDETVMGCILAKAHGEMLALAGAIAEVDEEALDEELLEMVSRSAEAAERKEK----- 324 Afu -TPETLAIDTEKVLEDFQRAYQMALNLAVNSAY-VTEETAEILIMKAVMDARNLAINAGI-FEKG--VMEDILMKAHAEMLSLASLLPDEA---LDEELKSMLSGIAEAAAQATPSV--- 308 Hma -EPEELELDIDEYRSDIQAAAGRAFNLSVNADY-PTATTAPTMLQSARGNAKSLALQAAI-EDPE--VVPDLVSKADAQVRALASQIDDEEA--LPEELQGVEADVATEEPTDDQDDDTA 316 Tac -SKEVLSLTPERIANDMAAAFSQAKAIAKSSMF-FVDEVLRDLLAEAKIKADALALAGQF-ITED--NVKDFLVRANVNAIILNNVLNKG----STQETPEEKKEEA------------- 294 Pto -NKDVLSISDEKIMSDLASAFLGAKSLAIEANF-IVPEIVPDMISKAYLEAQSLAIEANF-VDEK--NIDIFIRKAFINASAIEALINKD----LNNEDITDKKEEKA------------ 295 Pho -TPDVLAIDEQEYIDMLQKAYMHAFNLAVNIAY-PTPETIEAIIQKAFLNAKTVAIEAGY-ITKE--TIQDIIGRAFRAMLLLAQQLPEDV---LDEKTKELLSAQAQVAVATQPSEEE- 312 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Tma -NIAKL-PSKEELYAMLVGRVKAPITGLVFALSGIL-----------RNLVYVLNAI--KEKKSE------------ 179 Eco -RLATL-PTYEEAIARLMATMKEASAGKLVRTLAAV-----------------------RDAKEAA----------- 165 Sau -TVGSL-PSHDGLVSMLLSVLQAPVRNFAYAVKAIG-----------------------EQKEESAE---------- 166 Bsu -ALAEL-PPREGLLSMLLSVLKAPVRNLALAAKAVA-----------------------EQKEEQGA---------- 166 Smu -ALAAL-PNREGLLSMLLSVLQAPVRNVAYAVKAVA-----------------------ESKDEDAA---------- 167 Ljo -KLAAI-PGREGLLSMLVSVLQAPVRNFAYAVKAVA-----------------------DSKDE------------- 166 Mpu -AVAEL-PTYEEALTILASSLLGPLRQISLSFKLLV-----------------------DENKLSN----------- 168 Aae -ELAEL-PSKEELIAKLLGVLMAPVTQLAMTLKAVP-----------QKLVLVLKAI--EEEKSKGGQ--------- 199 Tth -ALAEL-PTMDELRAELVGVLQAPMAELVGVLGGVA-----------RELVGILEAYA-EKKAA------------- 173 Dra -RLASL-GSKTTLQAELVGVLSAHLSNFVGILEAYR-----------------------EKLEGEGGSESA------ 169 Cte -ALAAI-PPRDVLIAKLLGSFKAPVSNFAYLINAIK-----------------------EKKESEEA---------- 172 Fnu -ELAKL-PSREQLLSMLLNSMLGPIRKLAYATVAIA-----------------------DKKEGSAE---------- 170 Mle -RIADL-ESREVLLAKLAGAMKGTFAKAIGLFNAPT-----------SQMARLTAAL--QEKKAFEPASAE------ 177 Cdi -AIAEL-DNRETTLAKLAGAMKGNLAKAAGLFNAPA-----------SQVARLGAAL--QEKKEA------------ 171 San -KLADL-ESREVLLSKLAGAFKAKQSQAASVFQALP-----------SKLVRTVDAL--RAKQAEQGGAE------- 176 Nsp -ALGDL-PSKEQLMGQIAGAINALATKIAVGINEVP-----------GGLARALQAVA-DKENGGDDSAA------- 181 Gvi -AIADL-PPKEVLLAQFAGALKVLPTKIAVGINQVPTKVAVGINEVPAGLARVLEAL--RKQKEEQQQPQAAA---- 192 Lin -AIAGL-PSREQLLAQIAGGINGPARTIASGINQII-----------ASLARAIQATA-EKNNA------------- 177 Rpr -ELSKL-PSLHELRGKIVGLLQAPATKVVGVLQATS-----------SSIARVIHAHAIKH---------------- 169 Atu -SLATL-PSLDELRAKLLGLLNAPATRVATVVAAPA-----------SQLARVFSAYA-KKDEAA------------ 172 Hpy -AVSKL-PSKEELMGMLLSVWTAPARYFVTGLDNLR-----------------------KAKEEN------------ 164 Cje -ALAKM-PSRNELLAMLLQVWNAPITNFTIGLNALK-----------------------NKKESE------------ 159 Tpa -AYSKL-PGKKELVSMFLSALNATTVKFVRVLQAVM-----------------------DKRDEGVEVSVVSGGDSS 180 Bbu -AYSKL-PTKKESISLFASVLKAPVSKLARTLKALA-----------------------DVKN-------------- 162 Neu -ALAVL-PSREELLSKLLGTMQAPIAKFVRTLNEVP-----------SKFVRGLAAV--RDKK-------------- 171 Nme -ELASI-PSREELLSKLLFVMQAPVSGFARGLAALA-----------------------EKKAGEEAA--------- 166 Paer -VLASL-PTYDEAVSQLMSVIQGATSKLARTLAAIR-----------------------DQKEAAAA---------- 166 Bba -FLATL-PGKDQLRAMLLGTLLGAGSALARCLNAYA-----------------------EKMGGGAAAGTEEAPQA- 182

.......370.......380.......390.......400... Sce ------------EEAAAEEEEESDDDMGFGLFD---------- 312 Spo A----------------EEEEESDEDMGFGLFD---------- 312 Hsa KV------------EAKEESEESDEDMGFGLFD---------- 317 Dme -----------EAKKPESESEEEDDDMGFGLFD---------- 317 Cel ----------------EEPKEESDDDMGFGLFD---------- 312 Ddi ----------------EEKKEESDDDMGMGLFD---------- 305 Ath ---------------EEEAAEESDGDMGFDLFG---------- 317 Tcr AA------------EPEEEDDDDDFGMG-ALF----------- 322 Pfa V--------------EEEEEEEEDGFMGFGMFD---------- 316 Gla -------------EESDDDEIVGAGGM-FGGDSSSSEEESSSD 326 Bmo --------EKKEEKKEEKEEEESDDDMGFGLFD---------- 316 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Sac -----EKKEEKKEEEKKGPSEEEIGGGLSSLFGG--------- 335 Pae -------KEEAQEEKKEGPSEEEIAGSLASLF----------- 345 Ape -------------EEEEEKKEDQELSGLDSIFGGF-------- 344 Mva -------------KKEEKKEEAAPAAGLGLLF----------- 336 Mka EEEEEEEAEEEEAEEEEEEEEEEAAAGLGALFG---------- 357 Afu --EEREEEEKPEEEEEEEEKEEEAIEGLGALFG---------- 339 Hma SEDDADADDAAEEADDDDDDDEDAGDALGAMF----------- 348 Tac -------------KKEEKSPDESISEGLGALFQ---------- 314 Pto -------------EEKETSSDEDAASGLGALFG---------- 315 Pho ---KKEEEKTEEEEKEEEASEEEALAGLSALFG---------- 342