Supplementary Figure 1

Supplementary Figure 2

0.001 0.01 0.1 1 10 100-3

-2

-1

0

1

2

S score

GR

AV

Y sc

ore

0 0.01 0.1 1 10 100

Supplementary Figure 3

Reference Negative (RFU15s/min) +/-AtNMT1 247 13HsNMT1 269 17HsNMT2 281 7ScNMT 506 25PfNMT 206 15

Example (PfNMT: 206 ± 15)> 191 Red (-)

191-221 Yellow (+/-)>221 Green (+)

At1 Hs1 Hs2 Sc Pf At1 Hs1 Hs2 Sc PfA1 251 274 291 515 194 E1 228 235 271 505 192A2 855 438 490 1275 557 E2 281 327 300 477 190A3 1157 641 626 1328 364 E3 564 368 345 461 188A4 456 606 413 1773 356 E4 416 456 429 519 167A5 955 1404 1019 1570 392 E5 1760 1134 1195 1793 408A6 3309 1891 855 643 313 E6 725 630 969 741 250A7 1009 633 850 867 659 E7 603 991 710 1353 215A8 1311 483 818 839 341 E8 285 281 338 484 206A9 1007 607 926 1490 438 E9 559 776 968 474 219A10 865 772 1392 1895 662 E10 304 370 538 466 182A11 2561 1453 1316 2260 294 E11 261 294 333 452 211A12 1338 297 609 1477 530 E12 533 644 436 815 214B1 2571 1016 934 1838 555 F1 251 418 532 497 171B2 870 787 927 446 366 F2 321 262 287 467 179B3 2207 1093 1033 437 362 F3 235 258 377 388 176B4 2422 1196 964 1728 580 F4 283 300 326 427 220B5 537 855 623 456 272 F5 243 276 277 428 177B6 590 478 832 1345 354 F6 271 289 283 467 180B7 1569 809 1046 708 254 F7 242 194 279 476 187B8 754 491 489 1850 397 F8 345 363 354 433 201B9 1113 790 768 460 449 F9 232 139 269 453 185B10 1100 600 1002 472 291 F10 233 217 270 465 179B11 1386 745 932 478 190 F11 193 227 266 422 181B12 250 292 280 499 222 F12 243 258 277 518 221C1 245 275 271 471 186 G1 233 263 283 493 195C2 411 340 551 894 228 G2 230 246 267 476 185C3 1494 836 649 482 192 G3 215 216 271 447 182C4 1805 1140 1031 538 402 G4 180 231 269 457 190C5 1548 890 974 412 212 G5 195 114 273 452 157C6 1065 519 863 1658 401 G6 233 219 267 470 179C7 1274 894 1139 790 270 G7 210 216 261 465 182C8 1561 760 684 1976 251 G8 109 130 263 468 178C9 904 1130 1126 485 373 G9 209 211 271 295 177C10 1044 658 1087 452 192 G10 231 250 259 449 169C11 1179 340 855 430 208 G11 222 192 264 432 182C12 2074 3316 1523 945 380 G12 229 248 217 470 180D1 1630 935 1059 1447 207 H1 209 251 272 469 188D2 848 618 691 847 370 H2 215 243 273 456 178D3 1103 520 805 407 190 H3 208 210 266 530 168D4 352 522 479 472 184 H4 231 220 255 471 190D5 1418 914 1193 1276 450 H5 236 178 241 477 185D6 761 667 394 457 179 H6 219 269 253 456 171D7 1261 788 588 446 307 H7 225 216 271 501 190D8 875 481 684 460 274 H8 187 179 269 455 170D9 1122 907 985 1350 220 H9 173 215 245 509 165D10 1038 292 579 482 216 H10 213 209 267 419 188D11 1361 640 1232 659 216 H11 229 223 269 471 179D12 272 273 288 485 208 H12 250 282 285 554 223

Tecan InfiniteM200

Step 1Using a multichannel pipette a 96-well plate is manually loaded with 89 different peptides (5 µL of 2 mM stock solutions) and 7 negative control (5 µL of water) . The plate is then inserted in the fluorimeter (here a Tecan Infinite M200).

Microinjectors

A B

Step 2Microinjector A is filled with the reaction mixture (except MYR-CoA) and microinjector B with a solution of MYR-CoA 400µM.Once the paque is inserted in the fluorimeter, injector A injects 85 µL of reaction Mix per well.The 96-well plate is cooled at 30ºC for 3 min to stabilizate the reaction mixture.

Step 3Microinjector B injects 10 µL of MYR-CoA soution to star the in vitro reaction of MYR. Plate is automatically shaked horizontally to mix the reaction compounds. The reactions are ready to be monitored on the computer.

Supplementary Figure 4

Step 4After 10 min the reaction finishes and the fluorimeter software (Magellan) shows the maximum slope (RFU/min) according to the conditions fixed by the user. These values are obtained as output.

Step 5The values obtained as output data are converted with a spreadsheet software to generate a .xls file; the final data table is obtained.

1 2 3 4 5 6 7 8 9 10 11 12A 251 855 1157 456 955 3309 1009 1311 1007 865 2561 1338

B 2571 870 2207 2422 537 590 1569 754 1113 1100 1386 250

C 245 411 1494 1805 1548 1065 1274 1561 904 1044 1179 2074

D 1630 848 1103 352 1418 761 1261 875 1122 1038 1361 272

E 228 281 564 416 1760 725 603 285 559 304 261 533

F 251 321 235 283 243 271 242 345 232 233 193 243

G 233 230 215 180 195 233 210 109 209 231 222 229

I 209 215 208 231 236 219 225 187 173 213 229 250

Step 6Colour code is assigned according to the average of the values from negative reactions (A1,B12,C1…). Standard deviations are used to narrow the number of yellow boxes.

1 2 3 4 5 6 7 8 9 10 11 12A                        B                        A                        B                        A                        B                        A                        B