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% deuterium labeling 0 20 40 60 80 S-methyl propanethioate S-Methyl 2-methylbutanethioate S-methylbutanethioate S-methylthio acetate S-methyl 2-methylpropanethioate dimethyl trisulfide +3 dimethyl disulfide +3 dimethyl disulfide +6 PI PI 414723 dimethyl trisulfide +6 Dulce 'Dulce' ethyl (methylthio) acetate methyl 2-methylthio acetate 3(methylyhio) propanal ethyl 3(methylyhio) propionate methyl 3-methylthiopropionate 3(methylthio) propyl acetate 3 (methylthio) propanol Supplemental figure S1 Incorporation levels of 2H3-[methyl]-L-methionine into aroma volatiles in ripe melon fruit flesh. Bars represent enrichment values obtained in ‘Dulce’ (red) and ‘PI 414725’ (blue). Ripe melon flesh cubes were incubated with 5 mM 2H3-[methyl]-L-methionine for 6 hours, and the volatiles were analyzed by GC-MS. Levels are averages of 3 biological repeats ± SE. CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 1 1 1 1 1 1 1 1 MAELKNHN....FRGTKRASPDESDDVKTDSKKPTAMISSLLEDPAAAIANTRHEFGEHGGVNMSIEASATFTVMEPETM MAHFLETQEPLVFSGKKRNDRDDEDGDALVAKKSALAVCD..ADPAAAIANIRHEFGEHGGVNMSIEASATFTVMEPDTM ................MSITQNGIS....................TRSVHSGANPESHTGSVVAPIFQTSTFMMDTP... ............MRDSHNNTGFS......................TRAIHHGYDPLSHGGALVPPVYQTATYAFPTVEYG .............MTAQDIT.........................TTLLH..PKGDHVLHSHAYPIFQTSTFCFDSTQQG ............MSQLKDLQ.........................TRVLHTPSKWGEPLHAHTFPIFQTSTYLFDDTQMG .............MSHERMTPA.......................TACIHANPQKD.QFGAAIPPIYQTSTFVFDNCQQG .............MSGHAIDPTHTD....................TLSIHANPQKD.QFGAIVAPIYQTSTFLFDNCDQG CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 77 79 42 47 41 44 44 47 RRMFAGELGPDRDFFIYSRHFNPTVLNLSRQMAALEGTAAAYCTSSGMSAIAAVLLQLLASGDHVVASRTLYGGTHALMA RRMFTGELGPDNDFYVYSRHFNPTVLNLSRQMAALEGTQAAYCTSSGMSAISSVMLQLCSSGGHVVAASTLYGGTHALLS ....GQT....RAGFDYARTGTPNRSDLEDVLCELENASFAAAVNSGTSAEVAVFSALLGPGDEIIIPRDIYGGTYRLLK AACFAGE....EPGHFYSRISNPTLALLEQRMASLEGGEAGLALASGMGAITSTIWTLLRPGDELIVGRTLYGCTFSFLH ADLFMGK....GEGHIYSRLGNPTVEQFEEMVCSIEGAAGSAAFGSGMGAISSSTLAFLQKGDHLIAGDTLYGCTVSLFT ADLFGGK....REGHMYSRMGNPTVEYFEELVNSLEGGVGTAAFGSGMGAIHSATMGLLKAGDHLISGDTLYGCTVELFG GNRFAGQ....ESGYIYTRLGNPTVSNLEGKIAFLEKTEACVATSSGMGAIAATVLTILKAGDHLISDECLYGCTHALFE GARFGGK....EAGYMYTRIGNPTNSALEGKIAKLEHAEACAATASGMGAIAASVWTFLKAGDHLISDDCLYGCTHALFE CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 157 159 114 123 117 120 120 123 HFFPRTSNITTTFVDIGDLKEVEKAIVEGKTKVLYFESVSNPTLAVANIPELCRIGH.EKGVTVVVDNTFA.PMILSPAR HFLPRTCNITTSFVDITDHGAVANAIVEGRTQVLYFESVANPTLTVADIPELSRMAH.EKGVTVVVDNTFA.PMVLSPAK NEYERW.GISIRTVDLTDTEALAAAI.SAKTAIVWVETPSNPGLDIVDIAETAKLAH.AANAILAVDSTFATPILQRPIE HGIGEF.GVKIRHVDLNDAKALKAAI.NSKTRMIYFETPANPNMQLVDIAAVAEAVR.GHDVHVVVDNTYCTPYLQRPLE HWLPRF.GIEVDLIDTSDVEKVKAAW.KPNTKMVYLESPANPTCKVSDIKGIAVVCH.ERGARLVVDATFTSPCFLKPLE NRFKDM.GVDVSFVNTASDEAVVKAW.KPNTKMVYLESPANPTCKVSDIKGIAKICH.EKGAILVVDCTFTTPVFLPALE HALTKF.GIQVDFINTAIPGEVKKHM.KPNTKIVYFETPANPTLKIIDMERVCKDAHSQEGVLVIADNTFCSPMITNPVD HQLRKF.GVEVDFIDMAVPGNIEKHL.KPNTRIVYFETPANPTLKVIDIEDAVKQARKQKDILVIVDNTFASPILTNPLD CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 235 237 191 200 194 197 198 201 LGADVVVHSISKFISGGADIIAGAV.......CGP.TKLVNSMMDLRQGSLMLLGPTMNAKVAFELSERIPHLSLRMKEH LGADVVVHSISKFISGGADIIAGAV.......CGS.ENLVKEMMDLRGGSLMLLGPTMNAKVAFELSERIPHLGLRMREH LGADVVIHSTTKFINGHSDVIGGAVLAGDGSTCPRAAKVVERLE..S..YLASVGLGIAPFDAWLTRRGIKTLPVRMAKH LGADLVVHSATKYLSGHGDITAGLV.......VGR.KALVDRIR..LEGLKDMTGAVLSPHDASLLMRGIKTLALRMDRH LGADIALHSVSKYINGHGDVIGGVS.......SAKTAEDIATIK..F..YRKDAGSLMAPMDAFLCARGMKTLPIRMQIH LGADVVLHSITKYINGHGDVVGGVV.......TTKNPELLQKIK..A..FRKDTGSLMAPMDAFLCIRGVKTLPMRMKVH FGVDVVVHSATKYINGHTDVVAGLI.......CGK.ADLLQQIR..MVGIKDITGSVISPHDAWLITRGLSTLNIRMKAE LGVDIVVHSATKYINGHTDVVAGLV.......CSR.ADIIAKVK..SQGIKDITGAIISPHDAWLITRGTLTLDMRVKRA CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 307 309 267 270 263 266 268 271 CRRASVFAERMKKAG..LKVIYPGLEDHPQHQLMKSLANPEYGFGGMLCVDMG.TEERANKLMSILQNTTQFGFMAVSLG SHRAQVYAERMRDLG..MKVIYPGLETHPQHKLFKGMVNRDYGYGGLLSIDME.TEEKANKLMAYLQNATQFGFMAVSLG CENAQAVAQWLESRPEIAEVYYPGLPSHPGHEVAKKQMS...GFGGVVSFRTD.TEARALSLVKSTKLIT....LAESLG CANAQQVAEALARQPQVELIHYPGLPSFAQYELAQRQMR...LPGGMIAFELKGGIEAGRRFMNALQLFA....RAVSLG MENGLKVAKFLEQHEKIVKVNHPGLESFPGHDIAKKQMT...GYGSTFSFEMK.SFEAAKKLMEHLKVCT....LAVSLG MENGLKVAKFLEQHPKIKQVNHPGLESFPGHKIAMEQMK...GFGSTFSFEMK.SFEAAKKLMEHVKLCG....LAVSLG SENAMKVAEYLKSHPAVEKVYYPGFEDHEGHDIAKKQMR...MSGSMITFILKSGFEGAKKLLDNLKLIT....LAVSLG AENAQKVAEFLHEHKAVKKVYYPGLPDHPGHEIAKKQMK...MFGSMIAFDVD.GLEKAKKVLDNCHVVS....LAVSLG CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 384 386 339 343 335 338 341 343 YYETLMSCSGSSTSSEMSGEERELAGISPGLVRMSIGYMGTLEQKWSQFEKALAKVQDIGVPFCNN.............. YYETLMSCSGSSTSSELDPSQKEAAGISPGLVRMSVGYVGTLEQKWTQFEKAFLRM........................ GVESLIDHPATMTHLAVADCEL...SVSPTFIRLSLG.IEDIADILADLDAALASTVQTPDGVSSDAEALGAEVPIADPS DAESLAQHPASMTHSSYTAKERAHHGISEGLVRLSVG.LEDVEDLLADVEQALQACS....................... CVDTLIEHPASMTHAAVPENIMRKQGITPELVRISVG.IENVDDIIADLKQALELW........................ TLDTLIEHPASMTHAAVPEHLLKQQGLTRELVRISVG.LENPDDIIADLKQALEQC........................ GCESLIQHPASMTHAVVPKEEREAAGITDGMIRLSVG.IEDADELIADFKQGLDALL....................... GPESLIQHPASMTHAGVPKEEREAAGLTDNLIRLSVG.CENVQDIIDDLKQALDLVL....................... CmMGL AtMGL BlMGL PpMGL EhMGL1 EhMGL2 TvMGL1 TvMGL2 ........... ........... 415 PSTLQHPVATV ........... ........... ........... ........... ........... Supplemental figure S2 Supplemental figure S2. Multiple amino acid sequence alignment of CmMGL and other functionally characterized MGL's.. Alignments were performed using the EBI 'MUSCLE' tool and shaded using BoxShade version 3.21. Amino acids conserved in four genes or more are shaded in black. Amino acid similarities are shaded in gray. Red background indicates the conserved Lys residues that covalently bind the PLP cofactor and the pink background indicates conserved amino acids important for the formation of the homotetramer according to X-ray crystal analyses of PpMGL. CmMGL: Cucumis melo L-methionine-γ-lyase, GeneBank accession no. JX673982); AtMGL: Arabidopsis thaliana L-methionine γ-lyase, GeneBank accession no. AEE34271.1; BlMGL: Brevibacterium linens L-methionine-γ-lyase, GeneBank accession no. AAV54600; PpMGL: Pseudomonas Putida L-methionine-γ-lyase. GeneBank accession no. 1GC2_A; EhMGL1: Entamoeba histolytica L-methionine γ-lyase 1, GeneBank accession no. BAC75877; EhMGL2: Entamoeba histolytica L-methionine γ-lyase 2, GeneBank accession no. BAC75878; TvMGL1: Trichomonas vaginalis Lmethionine γ-lyase 1, GeneBank accession no. CAA04124; TvMGL2: Trichomonas vaginalis L-methionine γ-lyase 2, GeneBank accession no. CAA04125.