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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.