SUPPLEMENT 
 
Crystal structure of the N-terminal domains of the 
surface cellantigen4 of Rickettsia 
 
Jun Hyuck Lee,1,2 Clemens Vonrhein,3 Gerard Bricogne,3 andTina Izard1* 
 
1Cell Adhesion Laboratory,DepartmentofCancer Biology,TheScripps Research 
Institute,Jupiter,FL33458 
2Currentaddress:Division ofPolar LifeSciences,Korea Polar Research Institute
3Global PhasingLtd,Sheraton House,CastlePark,CambridgeCB3 0AX,England
 
*Correspondence to:T.Izard;E-mail:jholmes@scripps.edu
 
 
 
 
 
 
SupplementaryFigures
 
SupplementaryFigure S1.Multiplesequencealignmentofsca4 homologs from 
Rickettsia rickettsii (RI,UniProtKB codeB0BXR4),Rickettsia conorii(CO,UniProtKB 
codeQ52658),Rickettsia africae(AF,UniProtKB codeQ9AJ83),Rickettsia japonica 
(JA,UniProtKB codeQ9AJ79),Rickettsia felis(FE,UniProtKB codeQ9AJ37),and
Rickettsia prowazekii(PR,UniProtKB codeQ9ZD49).In thelinebelow thealignment, 
theasterisk (*) indicates thepositionof a single,fully conservedresidue(highlightedin 
yellow),thecolon (:) indicates conservation between groups ofstrongly similar 
properties,andtheperiod(.) indicates conservation between groups ofweakly similar 
properties accordingtoClustalX(http://www.ebi.ac.uk/Tools/msa/clustalw2/).The
secondary structural elements within thesca4 sequencefrom Rickettsia rickettsii 
predictedusingthePhyreserver (http://www.sbg.bio.ic.ac.uk/~phyre/) areshown above
thealignments. Notably, theseresults indicateda longflexibleloop (residues 1-20) at
theN-terminal region ofsca4. 2 
SupplementaryFigure S1 
                                         ---------------------h--hhhhhhhhhhhh
RI -------------------------------MRGFMSKDGNLDTSEFDPLANKEYTEEQKQTLEQEQ 36 
CO -----------------------------------MSKDGNLDTSEFDPLANKEYTEEQKQTLEQEQ 32 
AF -------------------------------MRGFMSKDGNLDTSEFDTLANKEYTAEQKQTLEQGQ 36 
JA -----------------------------------MSKDGNLNTSEFDPLANKEYTEEQKQTLEQEQ 32 
FE MSKDSDNPGYESGYESDTEEKKQEQAVPAQPISSTANKDGNPDTSEFDPLANKEYTEEQKQKLEQEQ 67 
PR -----------------------------------MSKNGNQDISEFDPLN-REFTEAEKQQQMQQE 31 
                                       .*:** : ****.*  :*:*  :**   * :
 
   hhhhh------------ssss----------hhh-------------hhhhhhhhhhh---hhhhhh
RI KEFLSQTTTPELEADDGFIVTSESSAQSTPSMSALSGNISPDSQTSDPITKAVRETIIQPQKDNLIE 103
CO KEFLSQTTTPALEADDGFIVTSASFAQSTPSMSALSGNISPDSQTSDPITKAVRETIIQPQKDNLIE 99
AF KEFLSQTTTPELEADDGFIVTSASFAQSTPSMSALSGNISPDSQTSDPITKAVRETIIQPQKDNLIE 103
JA KEFLSQTTTPELEADDGFIVTSASSAQSTPSTSALSGNISPDSQTSDPITKAVRETIIQPQKDNLIE 99
FE KEYFSQTTPQELEADDGFSFTPASSTQSTPSISSLSGGISSDSQTSDPITKAVRETIIQPQKDEIAE 134
PR QEFFSQTILD--IADDGFMVASSS--QATPSISFLSNNRPHGDHKSDPITEAIRKEILEKQRD----90
   :*::***      ***** .:. *  *:*** * **.. . ..:.*****:*:*: *:: *:*    
 
   hhhhhhhhh--hhhhhhhhhhhhhh--hhhhhhhh------hhhhhhhh--hhhhhhhhh-hhhhhh
RI QILKDLAALTDRDLAEQKRKEIEEEKEKDKTLSTFFGNPANREFIDKALEKPELKKKLESIEIAGYK170
CO QILKDLAALTDRDLAEQKRKEIEEEKEKDKTLSTFFGNPANREFIDKALENPELKKKLESIEIAGYK166
AF QILKDLAALTDRDLAEQKRKEIEEEKEKDKTLSTFFGNPANREFIDKALDNPELKKKLESIEIAGYK170
JA QILKDLAALTDHDLAEQKRKEIEEEKDKDKTLSTFFGNPANREFIDKALENPELKKKLESIEIAGYK166
FE QILKDLAALADRDLAEQKRKEIEEE--KDKTLSAFFGNPANREFIDKALENPELKKKLESIEIAGYK199
PR -ILREYFVNTNPELAEQIAK--EED---DRKFRAFLSNQDNYALINKAFEDTKTKKNLEKAEIVGYK151
    **::  . :: :****  *  **:   *:.: :*:.*  *  :*:**::..: **:**. **.*** 
 
   hhhhhhhhh-----------shhhss-----sssss-----hhhhh---------ssssss---sss
RI NVHNTFSAASGYPGGFKPVQWENHVSASDLRATVVKNDAGDELCTLNETTVKTKPFTLAKQDGTQVQ237
CO NVHNTFSAASGYPGGFKPVQWENHVSANDLRATVVKNDAGDELCTLNETTVKTKPFTLAKQDGTQVQ233
AF NVHNTFSAASGYPGGFKPVQWENHVSASDLRATVVKNDAGDELCTLNETTVKTKPFTLAKQDGTQVQ237
JA NVHNTFSAASGYPGGFKPVQWENHVSASDLRATVVKNDAGDELCTLNETTVKTKPFTLAKQDGTQVQ233
FE NVLSTYSAANGYQGGFKPVQWENQISASDLRATVVRNDAGDELCTLNETTVKTKPFTVAKQDGTQVQ266
PR NVLSTYSVANGYQGGFQPVQWENQVSASDLRSTVVKNDEGEELCTLNETTVKTKDLIVAKQDGTQVQ218
   ** .*:*.*.** ***:******::**.***:***:** *:************* : :*********
 
   sssssss---hh--------ssssssss-----------sssssss----------ss---------
RI ISSYREIDFPIKLDQADGSMHLSMVALKADGTKPSKDKAVYFTAHYEEGPNGKPQLKEISSPKPLKF304
CO ISSYREIDFPIKLDKADGSMHLSMVALKADGTKPSKDKAVYFTAHYEEGPNGKPQLKEISSPKPLKF300
AF ISSYREIDFPIKLDKADGSMHLSMVALKADGTKPSKDKAVYFTAHYEEGPNGKPQLKEISSPKPLKF304
JA ISSYREIDFPIKLDKADGSMHLSMVALKADGTKPSKDKAVYFTAHYEEGPNGKPQLKEISSPKPLKF300
FE INSYREIDFPIKLDKADGSMHLSMVALKADGTKPSKDKAVYFTAHYEEGPNGKPQLKEISSPKPLKF333
PR INSYREINFPIKLDKANGSMHLSMVALKADGTKPAKDKAVYFTAHYEEGPNGKPQLKEISSPQPLKF285
   *.*****:******:*:*****************:***************************:****
 
   --------ssss---ssssss----hhhhhhhhhhh--------hh--hhhhhhh-------------
RI AGTGDDAIAYIEHGGEIYTLAVTRGKYKEMMKEVELNQGQSVDLSQ--AEDIIIGQGQSKE--QPLI 367
CO AGTGDDAIAYIEHGGEIYTLAVTRGKYKEMMKEVELNQGQSVDLSQ--AEDIIIGQGQSKE--QPLI 363
AF AGTGDDAIAYIEHGGEIYTLAVTRGKYKEMMKEVELNQGQSVDLSQ--AEDIIIGQGQSKE--QPLI 367
JA AGTGDDAIAYIEHGGEIYTLAVTRGKYKEMMKEVELNQGQSVDLSQ--AEDIIIGQGQSKE--QPLI 363
FE AGDGPDAVAYIEHGGEIYTLAVTRGKYKEMMREVELNQGQSVDLSQTIAEDLTKVQGRSQETPQPII 400
PR VGTGDDAVAYIEHGGEIYTLAVTRGKYKEMMKEVALNHGQSVALSQTIAEDLTHVQGPSHETHKPII 352
   .* * **:***********************:****:**** ***  ***:   ** *:*  :*:* 3 
 
 
 
SupplementaryFigure S2
 
 
 
Interdomaininterface of sca4 (residues 21-360)from Rickettsia rickettsii
 
 
(A)Hydrophobic interactions 
 
N-terminaldomain C-terminaldomain Distance 
Ile-85 CG2   Phe-186 CZ   3.8 Å
Leu-101 CD2  Leu-348 CD2  3.9 Å
Phe-139 CD1  Val-346 CG1  4.3 Å
Leu-152 CD2  Pro-301 CB   4.2 Å
 
 
 
(B)Polar interactions 
 
N-terminaldomain C-terminaldomain Distance 
Arg-90 NH2  Lys-294 O   3.7 Å
Asp-83 OD2  His-258 NE2  3.3 Å
Arg-90 NH2  Glu-295 OE1  2.9 Å 
Lys-98 NZ   Gln-344 O   2.8 Å
Glu-153 O   Gln-231 NE2  2.9 Å
Lys-158 NZ   Glu-316 O   2.6 Å
Tyr-169 OH   Ser-297 OG   2.8 Å
Lys-170 NZ   Asp-274 OD1  3.9 Å