Page 1 1:5,000 1: 10,000 1: 25,000 1: 100,000 ...

Supplementary Fig. S1 A

1: 5,000 1: 10,000

1: 25,000 1: 100,000

F T C M C T F

F T C M C T F

kDa 250 130 100

55

35

1

B

α CDPK rabbit 1, dilution series B

10

y = 0.0617x - 0.1739 R² = 0.9994

9 Cell number x 106

8 7

6

Cell # x 10

Linear (Series1)

9 3 1 0.3

5 4

6

Series1

kDa

3

250

2

130

1

100

0 0

50

100 Signal strength

150

200

055 35

15

2

C Tg kDa

5

Sn-e 1

2 0.4

Sn-i BT 0.3 0.3

250 130 100 55 35

3

Supplementary Fig. S2 A gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

---MGQQESTLG-----------GAAGEPRSRGHAAGTSGGPGDHLHATPGMFVQHSTAI ---MGQEQSTLASTSPTAAHQSSGPGGERLRSGGGTAGGGGADDKLHATPGMFVQHSTAV MSAAGDRASPY-VSAPLASQKSLRP-------STDVETHRETPVCVSLSPSLCTYTHMYT *:. * . . : :*.: .

46 57 52

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

FSDRYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQL FSDRYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQL YVRGYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQL : ********************************************************

106 117 112

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

DHPNIMKLYEFFEDKGYFYLVGEVYTGGELFDEIISRKRFSEVDAARIIRQVLSGITYMH DHPNIMKLYEIFEDRGYYYLVGEVYCGGELFDEIISRKRFSEVDAARIIRQVLSGITYMH DHPNIMKLYEIFEDRGYYYLVGEVYCGGELFDEIISRKRFSEVDAARIIRQVLSGITYMH **********:***:**:******* **********************************

166 177 172

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

KNKIVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGTAYYIAPEVLHGT KNKVVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGTAYYIAPEVLHGT KNKVVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGTAYYIAPEVLHGT ***:********************************************************

226 237 232

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

YDEKCDVWSTGVILYILLSGCPPFNGANEYDILKKVEKGKYTFELPQWKKVSESAKDLIR YDEKCDVWSTGVILYILLSGCPPFNGADEYEILKKVEKGKYTFELPQWKKVSESAKDLIR YDEKCDVWSTGVILYILLSGCPPFNGADEYEILKKVEKGKYTFELPQWKKVSESAKDLIR ***************************:**:*****************************

286 297 292

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

KMLTYVPSMRISARDALDHEWIQTYTKEQISVDVPSLDNAILNIRQFQGTQKLAQAALLY KMLTYVPSMRISARDALDHEWVQTYGKEQATVDVPSLDNAILNIRQFQGTQKLSQAALLY KMLTYVPSMRISARDALDHEWVQTYGKEQATVDVPSLDNAILNIRQFQGTQKLSQAALLY *********************:*** *** :**********************:******

346 357 352

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

MGSKLTSQDETKELTAIFHKMDKNGDGQLDRAELIEGYKELMRMKGQDASMLDASAVEHE MGSKLTSKEETRELSAIFHRMDKNGDGQLDRAELIEGYKELLRLKGQDASLLDQSAVEYE MGSKLTSKEETRELSAIFHRMDKNGDGQLDRAELIEGYKELLRLKGQDASLLDQSAVEYE *******::**:**:****:*********************:*:******:** ****:*

406 417 412

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

VDQVLDAVDFDKNGYIEYSEFVTVAMDRKTLLSRERLERAFRMFDSDNSGKISSTELATI VDQVLEAVDFDKNGFIGYSEFVTVAMDRKSLLSRERLERAFKMFDADNSGKISSAELATI VDQVLEAVDFDKNGFIGYSEFVTVAMDRKSLLSRERLERAFKMFDADNSGKISSAELATI *****:********:* ************:***********:***:********:*****

466 477 472

gi|12484153|gb|AAG53993.1|AF333958_1 SN3_02800495 SRCN_3314_SN1

FGVSDVDSETWKSVLSEVDKNNDGEVDFDEFQQMLLKLCGN-----FGVSDIDSETWKAVLAEVDRNNDGEVDFGEFQEMLLKLCDENATKKR FGVSDIDSETWKAVLAEVDRNNDGEVDFGEFQEMLLKLCDENATKKR *****:******:**:***:******** ***:****** :

507 524 519

4

B

SnCDPK1 TGME49_CDPK1

MGQEQSTLASTSPTAAHQSSGPGGERLRSGGGTAGGGGADDKLHATPGMFVQHSTAVFSD MGQQESTLG-----------GAAGEPRSRGHAAGTSGGPGDHLHATPGMFVQHSTAIFSD ***::***. * .** * .:. .** *:**************:***

60 49

SnCDPK1 TGME49_CDPK1

RYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQLDHP RYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQLDHP ************************************************************

120 109

SnCDPK1 TGME49_CDPK1

NIMKLYEIFEDRGYYYLVGEVYCGGELFDEIISRKRFSEVDAARIIRQVLSGITYMHKNK NIMKLYEFFEDKGYFYLVGEVYTGGELFDEIISRKRFSEVDAARIIRQVLSGITYMHKNK *******:***:**:******* *************************************

180 169

SnCDPK1 TGME49_CDPK1

VVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGTAYYIAPEVLHGTYDE IVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGTAYYIAPEVLHGTYDE :***********************************************************

240 229

SnCDPK1 TGME49_CDPK1

KCDVWSTGVILYILLS-------------------------------------------KCDVWSTGVILYILLSGESVPRSAFPVSPRSSSSFCSSSSFCSPGLPVYLQKREIKSCVA ****************

256 289

SnCDPK1 TGME49_CDPK1

-------------------------------GCPPFNGADEYEILKKVEKGKYTFELPQW FYVGMQRDACALIYLGTQDARAGAFFPDEKPRCPPFNGANEYDILKKVEKGKYTFELPQW *******:**:*****************

285 349

SnCDPK1 TGME49_CDPK1

KKVSESAKDLIRKMLTYVPSMRISARDALDHEWVQTYGKEQATVDVPSLDNAILNIRQFQ KKVSESAKDLIRKMLTYVPSMRISARDALDHEWIQTYTKEQISVDVPSLDNAILNIRQFQ *********************************:*** *** :*****************

345 409

SnCDPK1 TGME49_CDPK1

GTQKLSQAALLYMGSKLTSKEETRELSAIFHRMDKNGDGQLDRAELIEGYKELLRLKGQD GTQKLAQAALLYMGSKLTSQDETKELTAIFHKMDKNGDGQLDRAELIEGYKELMRMKGQD *****:*************::**:**:****:*********************:*:****

405 469

SnCDPK1 TGME49_CDPK1

ASLLDQSAVEYEVDQVLEAVDFDKNGFIGYSEFVTVAMDRKSLLSRERLERAFKMFDADN ASMLDASAVEHEVDQVLDAVDFDKNGYIEYSEFVTVAMDRKTLLSRERLERAFRMFDSDN **:** ****:******:********:* ************:***********:***:**

465 529

SnCDPK1 TGME49_CDPK1

SGKISSAELATIFGVSDIDSETWKAVLAEVDRNNDGEVDFGEFQEMLLKLCDENATKKR* SGKISSTELATIFGVSDVDSETWKSVLSEVDKNNDGEVDFDEFQQMLLKLCGN*-----******:**********:******:**:***:******** ***:****** :

524 582

5

C Sn3 TgCDPK1

MGQEQSTLASTSPTAAHQSSGPGGERLRSGGGTAGGGGADDKLHATPGMFVQHSTAVFSD MGQQESTLG-----------GAAGEPRSRGHAAGTSGGPGDHLHATPGMFVQHSTAIFSD ***::***. * .** * .:. .** *:**************:***

60 49

Sn3 TgCDPK1

RYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQLDHP RYKGQRVLGKGSFGEVILCKDKITGQECAVKVISKRQVKQKTDKESLLREVQLLKQLDHP ************************************************************

120 109

Sn3 TgCDPK1

NIMKLYEIFEDRGYYYLVGEVYCGGELFDEIISRKRFSEVDAARIIRQVLSGITYMHKNK NIMKLYEFFEDKGYFYLVGEVYTGGELFDEIISRKRFSEVDAARIIRQVLSGITYMHKNK *******:***:**:******* *************************************

180 169

Sn3 TgCDPK1

VVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGTNGPHFAAAVGVAAAA IVHRDLKPENLLLESKSKDANIRIIDFGLSTHFEASKKMKDKIGT--------------:********************************************

240 214

Sn3 TgCDPK1

AAIRVAAAVGVVATAVGLAAAAVDVAATVDVASAALGVVLLVLLETERPWAYYIAPEVLH --------------------------------------------------AYYIAPEVLH **********

300 224

Sn3 TgCDPK1

GTYDEKCDVWSTGVILYILLS--------------------------------------GTYDEKCDVWSTGVILYILLSGESVPRSAFPVSPRSSSSFCSSSSFCSPGLPVYLQKREI *********************

321 284

Sn3 TgCDPK1

------------------------------------GCPPFNGADEYEILKKVEKGKYTF KSCVAFYVGMQRDACALIYLGTQDARAGAFFPDEKPRCPPFNGANEYDILKKVEKGKYTF *******:**:************

345 344

Sn3 TgCDPK1

ELPQWKKVSESAKDLIRKMLTYVPSMRISARDALDHEWVQTYGKEQATVDVPSLDNAILN ELPQWKKVSESAKDLIRKMLTYVPSMRISARDALDHEWIQTYTKEQISVDVPSLDNAILN **************************************:*** *** :************

405 404

Sn3 TgCDPK1

IRQFQGTQKLSQAALLYMGSKLTSKEETRELSAIFHRMDKNGDGQLDRAELIEGYKELLR IRQFQGTQKLAQAALLYMGSKLTSQDETKELTAIFHKMDKNGDGQLDRAELIEGYKELMR **********:*************::**:**:****:*********************:*

465 464

Sn3 TgCDPK1

LKGQDASLLDQSAVEYEVDQVLEAVDFDKNGFIGYSEFVTVAMDRKSLLSRERLERAFKM MKGQDASMLDASAVEHEVDQVLDAVDFDKNGYIEYSEFVTVAMDRKTLLSRERLERAFRM :******:** ****:******:********:* ************:***********:*

525 524

Sn3 TgCDPK1

FDADNSGKISSAELATIFGVSDIDSETWKAVLAEVDRNNDGEVDFGEFQEMLLKLCDENA FDSDNSGKISSTELATIFGVSDVDSETWKSVLSEVDKNNDGEVDFDEFQQMLLKLCGN-**:********:**********:******:**:***:******** ***:****** :

585 582

Sn3 TgCDPK1

TKKR ----

589 582

6