Figure S1. Sequence alignment of a hemoglobin alpha subunit ...

Figure S1. Sequence alignment of a hemoglobin alpha subunit. Tyrosine C7 (Tyr 42, human) is highly conserved. P69905 P69907 P01958 P01965 P01966 P01945 P01994 P83135 P01999 P02018 Q1AGS9 P14522 P07408

MVLSPADKTNVKAAWGKVGAHAGEYGAEALERMFLSFPTTKTYFPHF-DLSHGSAQVKGH MVLSPADKTNVKAAWGKVGAHAGEYGAEALERMFLSFPTTKTYFPHF-DLSHGSAQVKGH MVLSAADKTNVKAAWSKVGGHAGEYGAEALERMFLGFPTTKTYFPHF-DLSHGSAQVKAH -VLSAADKANVKAAWGKVGGQAGAHGAEALERMFLGFPTTKTYFPHF-NLSHGSDQVKAH MVLSAADKGNVKAAWGKVGGHAAEYGAEALERMFLSFPTTKTYFPHF-DLSHGSAQVKGH -VLSAKDKTNISEAWGKIGGHAGEYGAEALERMFFVYPTTKTYFPHF-DVSHGSAQVKGH MVLSAADKNNVKGIFTKIAGHAEEYGAETLERMFTTYPPTKTYFPHF-DLSHGSAQIKGH MVLTAGDKANVKTVWSKVGSHLEEYGSETLERLFIVYPSTKTYFPHF-DLHHDSAQVRAH MVLSMEDKSNVKAIWGKASGHLEEYGAEALERMFCAYPQTKIYFPHF-DMSHNSAQIRAH -SLSDKDKAVVKALWAKIGSRADEIGAEALGRMLTVYPQTKTYFSHWSDLSPGSGPVKKH MSLSAKDKATVKDFFGKMSTRSDDIGAEALSRLVAVYPQTKSYFAHWKSASPGSAPVRKH -VLSAADKTNVKSAFSKIGGQADEYGAETLERMFATYPQTKTYFPHF-DLGKGSAQVKAH -VLSAADKTAIKHLTGSLRTNAEAWGAESLARMFATTPSTKTYFSKFTDFSANGKRVKAH *: ** :. . . *:*:* *:. * ** **.:: . .. :: *

59 59 59 58 59 58 59 59 59 59 60 58 59

P69905 P69907 P01958 P01965 P01966 P01945 P01994 P83135 P01999 P02018 Q1AGS9 P14522 P07408

GKKVADALTNAVAHVDDMPNALSALSDLHAHKLRVDPVNFKLLSHCLLVTLAAHLPAEFT GKKVADALTNAVAHVDDMPNALSALSDLHAHKLRVDPVNFKLLSHCLLVTLAAHLPAEFT GKKVGDALTLAVGHLDDLPGALSNLSDLHAHKLRVDPVNFKLLSHCLLSTLAVHLPNDFT GQKVADALTKAVGHLDDLPGALSALSDLHAHKLRVDPVNFKLLSHCLLVTLAAHHPDDFN GAKVAAALTKAVEHLDDLPGALSELSDLHAHKLRVDPVNFKLLSHSLLVTLASHLPSDFT GKKVADALTNAVGHLDDLPGALSALSDLHAHKLRVDPVNFKLLSHCLLVTLANHHPADFT GKKVVAALIEAANHIDDIAGTLSKLSDLHAHKLRVDPVNFKLLGQCFLVVVAIHHPAALT GRKVLSALGEAVNHIDDIPGALSKLSDLHAQTLRVDPVNFKLLNLCFVVVVGRHHPTILT GKKVFSALHEAVNHIDDLPGALCRLSELHAHSLRVDPVNFKFLAHCVLVVFAIHHPSALS GKTIMGAVGDAVSKIDDLVGALSALSELHAFKLRIDPANFKILAHNVIVVIGMLFPGDFT GITIMGGVYDAVGKIDDLKAGLLSLSELHAFMLRVDPVNFKLLAHCMLVCMSMVFPEEFT GKKVAAALVEAANAVDDIAGALSKLSDLHAQKLRVDPVNFKLLGQCFLVTVATHNPSLLT GGKVLNAVADATDHLDNVAGHLDPLAVLHGTTLCVDPHNFPLLTQCILVTLAAHL-TELK * .: .: *. :*:: * *: **. * :** ** :* .: .. :.

119 119 119 118 119 118 119 119 119 119 120 118 118

P69905 P69907 P01958 P01965 P01966 P01945 P01994 P83135 P01999 P02018 Q1AGS9 P14522 P07408

PAVHASLDKFLASVSTVLTSKYR PAVHASLDKFLASVSTVLTSKYR PAVHASLDKFLSSVSTVLTSKYR PSVHASLDKFLANVSTVLTSKYR PAVHASLDKFLANVSTVLTSKYR PAVHASLDKFFASVSTVLTSKYR PEVHASLDKFLCAVGTVLTAKYR PEVHVSLDKFLSAVATALTSKYR PEIHASLDKFLCAVSAVLTSKYR PEVHMSVDKFFQNLALALSEKYR PQVHVAVDKFLAQLALALCEKYR PEVHASLDKFLCAVGTVLTAKYR PETHCALDKFLCEVATALGSHYR * * ::***: :. .* :**

P69905 P69907 P01958 P01965 P01966 P01945 P01994 P14522 P83135 P01999 P02018 Q1AGS9 P07408

HBA_HUMAN HBA_PANTR HBA_HORSE HBA_PIG HBA_BOVIN HBA_MESAU HBA_CHICK HBA_TURME HBAA_GEONI HBA_ALLMI HBA_CARAU HBA1_BORSA HBA_SQUAC

142 142 142 141 142 141 142 142 142 142 143 141 141

Homo sapiens (Human) Hemoglobin alpha chain Pan troglodytes (Chimpanzee) Hemoglobin alpha chain Equus caballus (Horse) Hemoglobin alpha chain Sus scrofa (Pig) Hemoglobin alpha chain Bos taurus (Bovine) Hemoglobin alpha chain Mesocricetus auratus (Golden hamster) Hemoglobin alpha chain Gallus gallus (Chicken) Hemoglobin alpha-A chain Turdus merula (Blackbird) Hemoglobin alpha-A chain Geochelone nigra (Galapagos giant tortoise) Hemoglobin alpha-A chain Alligator mississippiensis (American alligator) Hemoglobin alpha chain Carassius auratus (Goldfish) Hemoglobin alpha chain Boreogadus saida (Polar cod) Hemoglobin alpha-1 chain Squalus acanthias (Spiny dogfish) Hemoglobin alpha chain

Figure S2. Sequence alignment of a hemoglobin beta subunit. Phenyalanine C7 (Phe 41, human) is largely conserved throughout mammals and birds, but is a tyrosine in amphibians and fish, whilst split between tyrosine and phenylalanine in reptiles. P68871 P68873 P02062 P02067 P02070 P02094 P02112 P14524 P83123 P02130 P02140 Q1AGS7 P07409

MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPK MVHLTPEEKSAVTALWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPK -VQLSGEEKAAVLALWDKVNEEEVGGEALGRLLVVYPWTQRFFDSFGDLSNPGAVMGNPK MVHLSAEEKEAVLGLWGKVNVDEVGGEALGRLLVVYPWTQRFFESFGDLSNADAVMGNPK --MLTAEEKAAVTAFWGKVKVDEVGGEALGRLLVVYPWTQRFFESFGDLSTADAVMNNPK MVHLTDAEKALVTGLWGKVNADAVGAEALGRLLVVYPWTQRFFEHFGDLSSASAVMNNPQ MVHWTAEEKQLITGLWGKVNVAECGAEALARLLIVYPWTQRFFASFGNLSSPTAILGNPM -VQWTAEEKQLITGLWGKVNVAECGGEALARLLIVYPWTQRFFASFGNLSSPTAVLGNPK MVHWTPEEKQYITSLWAKVNVEEVGGEALARLLIVYPWTQRFFSSFGNLSSASAILHNAK -ASFDAHERKFIVDLWAKVDVAQCGADALSRMLIVYPWKRRYFEHFGKMCNAHDILHNSK -VEWTDAERSAIIGLWGKLNPDELGPQALARCLIVYPWTQRYFATFGNLSSPAAIMGNPK MVEWTATERTHIEAIWSKIDIDVCGPLALQRCLIVYPWTQRYFGSFGDLSTDAAIVGNPK -VHWTGEEKALVNAVWTKTDHQAVVAKALERLFVVYPWTKTYFVKFNGKFHAS----DST *: : .* * . ** * ::****.: :* *. :.

60 60 59 60 58 60 60 59 60 59 59 60 55

P68871 P68873 P02062 P02067 P02070 P02094 P02112 P14524 P83123 P02130 P02140 Q1AGS7 P07409

VKAHGKKVLGAFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFG VKAHGKKVLGAFSDGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFG VKAHGKKVLHSFGEGVHHLDNLKGTFAALSELHCDKLHVDPENFRLLGNVLVVVLARHFG VKAHGKKVLQSFSDGLKHLDNLKGTFAKLSELHCDQLHVDPENFRLLGNVIVVVLARRLG VKAHGKKVLDSFSNGMKHLDDLKGTFAALSELHCDKLHVDPENFKLLGNVLVVVLARNFG VKAHGKKVIHSFADGLKHLDNLKGAFSSLSELHCDKLHVDPENFKLLGNMIIIVLSHDLG VRAHGKKVLTSFGDAVKNLDNIKNTFSQLSELHCDKLHVDPENFRLLGDILIIVLAAHFS VQAHGKKVLTSFGEAVKNLDSIKGTFAQLSELHCDKLHVDPENFRLLGDILVVVLAAHFG VLAHGKKVLTSFGDAVKNLDNIKKTFAQLSELHCEKLHVDPENFKLLGNILIIVLATRFP VQEHGKKVLASFGEAVKHLDNIKGHFANLSKLHCEKFHVDPENFKLLGDIIIIVLAAHHP VAAHGRTVMGGLERAIKNMDNIKATYAPLSVMHSEKLHVDPDNFRLLADCITVCAAMKFG VANHGVVALTGLRTALDHMDDIKATYATLSVLHSEKLHVDPDNFRLLCDCLTIVVAGKFG VQTHAGKVVSALTVAYNHIDDVKPHFVELSKKHYEELHVDPENFKLLANCLEVELGHALH * *. .: .: . ::*.:* : ** * :::****:**:** : : .

120 120 119 120 118 120 120 119 120 119 119 120 115

P68871 P68873 P02062 P02067 P02070 P02094 P02112 P14524 P83123 P02130 P02140 Q1AGS7 P07409

KE-FTPPVQAAYQKVVAGVANALAHKYH KE-FTPPVQAAYQKVVAGVANALAHKYH KD-FTPELQASYQKVVAGVANALAHKYH HD-FNPNVQAAFQKVVAGVANALAHKYH KE-FTPVLQADFQKVVAGVANALAHRYH KD-FTPSAQSAFHKVVAGVANALAHKYH KD-FTPECQAAWQKLVRVVAHALARKYH KD-FTPDCQAAWQKLVRVVAHALARKYH KE-FTPASQAAWTKLVNAVAHALALGYH ED-FSVECHAAFQKLVRQVAAALAAEYH PSGFNADVQEAWQKFLSVVVSALCRQYH PT-LRPEMQAAWQKYLSAVVSALGRQYH KE-FTPEVQAAWSKFSNVVVDALSKGYH : : : * *. ** **

P68871 P68873 P02062 P02067 P02070 P02094 P02112 P14524 P83123 P02130 P02140 Q1AGS7 P07409

HBB_HUMAN HBB_PANTR HBB_HORSE HBB_PIG HBB_BOVIN HBB_MESAU HBB_CHICK HBB_TURME HBB_GEONI HBB_ALLMI HBB_CARAU HBB1_BORSA HBB_SQUAC

147 147 146 147 145 147 147 146 147 146 147 147 142

Homo sapiens (Human) Hemoglobin beta chain Pan troglodytes (Chimpanzee) Hemoglobin beta chain Equus caballus (Horse) Hemoglobin beta chain Sus scrofa (Pig) Hemoglobin beta chain Bos taurus (Bovine) Hemoglobin beta chain Mesocricetus auratus (Golden hamster) Hemoglobin beta chain Gallus gallus (Chicken) Hemoglobin beta chain Turdus merula (Blackbird) Hemoglobin beta chain Geochelone nigra (Galapagos giant tortoise) Hemoglobin beta chain Alligator mississippiensis (American alligator) Hemoglobin beta chain Carassius auratus (Goldfish) Hemoglobin beta chain Boreogadus saida (Polar cod) Hemoglobin beta-1 chain Squalus acanthias (Spiny dogfish) Hemoglobin beta chain