Mith_00200 Mith_00200   Mith_00220 Mith_00220

Mith_00210 : CDS information

close this sectionLocation

Organism
StrainATCC 12956
Entry nameMithramycin
Contig
Start / Stop / Direction24,436 / 23,171 / - [in whole cluster]
24,436 / 23,171 / - [in contig]
Locationcomplement(23171..24436) [in whole cluster]
complement(23171..24436) [in contig]
TypeCDS
Length1,266 bp (421 aa)
Click on the icon to see Genetic map.

close this sectionAnnotation

Category2.3 modification addition of sugar moiety
ProductNADPH-dependent dTDP-4-keto-2,6-dideoxy-D-glucose C-4 ketoreductase/SAM-dependent dTDP-4-keto-2,6-dideoxy-D-glucose C-3 C-methyltransferase
NADPH-dependent dTDP-4-keto-D-olivose C-4 ketoreductase/SAM-dependent dTDP-4-keto-D-olivose C-3 C-methyltransferase
Product (GenBank)D-mycarose 3-C-methyltransferase
Gene
Gene (GenBank)mtmC
EC number
Keyword
  • D-olivose
  • D-mycarose
Note
  • bifunctional protein
Note (GenBank)
Reference
ACC
PmId
[11254130] The mtmVUC genes of the mithramycin gene cluster in Streptomyces argillaceus are involved in the biosynthesis of the sugar moieties. (Mol Gen Genet. , 2001)
[11853433] Ketopremithramycins and ketomithramycins, four new aureolic acid-type compounds obtained upon inactivation of two genes involved in the biosynthesis of the deoxysugar moieties of the antitumor drug mithramycin by Streptomyces argillaceus, reveal novel insights into post-PKS tailoring steps of the mithramycin biosynthetic pathway. (J Am Chem Soc. , 2002)
[21960454] Investigating Mithramycin deoxysugar biosynthesis: enzymatic total synthesis of TDP-D-olivose. (Chembiochem. , 2011)
[22997042] Cooperation of two bifunctional enzymes in the biosynthesis and attachment of deoxysugars of the antitumor antibiotic mithramycin. (Angew Chem Int Ed Engl. , 2012)
[25587924] Structural insight into MtmC, a bifunctional ketoreductase-methyltransferase involved in the assembly of the mithramycin trisaccharide chain. (Biochemistry. , 2015)
comment
[PMID:11254130](2001)
Streptornyces argillaceus ATCC 12956 cosmid clone cosAR3から3852bpのDNA領域をシークエンスしたところ、mtmV, mtmU, mtmCの3ORFsが見つかった。

mtmCの推定産物 (1266bp, 421aa, Mr45796)は、配列解析からD-mycaroseに関与するC-3 methyltransferaseをコードすると想定された。

mtmC不活化mutant産物はどれもD-mycaroseを含まず機能提唱に一致するが、さらにlower saccharide鎖D-olivose部分のC-4でketo基を含んでいる。これがpolar effectsのせいではないことが確認されたため、mtmCは3-C-methyltransferase活性に加えて、D-olivose生合成でのC-4 keto基の還元を担うretoreductase活性をコードしうると示唆される。

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[PMID:11853433](2002)
mtmC and mtmTIII の役割を調査。
mtmCについては[PMID:11254130]と同じ実験内容。産物の構造解析がより詳細。

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[PMID:21960454](2011)
dTDP-D-olivoseの酵素的全合成。
recombinant protein MtmCを用いて、in vitroでの反応を見ている。
dTDP-4-keto-2,6-dideoxy-D-glucose (=dTDP-4-keto-D-olivose) + MtmC + NADPH からdTDP-D-olivoseの形成が確認された。

よって、MtmCはD-mycarose生合成のためのC-methyltransferに加えて、D-olivose生合成のために必要な4-ketoreductionを実施できるbifunctional enzymeである。

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[PMID:22997042](2012)
MtmCの実験内容は[PMID:21960454]と同様。

さらに、MtmGIV, MtmC, MtmTIII とcofactorを組み合わせて糖からの生成物を確認。MtmCは、dTDP-4-keto-D-olivoseから、その4-ketoreductase and C-methyltransferase活性を通してD-olivose and D-mycaroseの両方の平行した形成を供給できることが明らかに示された。

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[PMID: 25587924](2015) abstract
MtmCを含む複合体の結晶構造解析。

close this sectionSequence

selected fasta
>NADPH-dependent dTDP-4-keto-2,6-dideoxy-D-glucose C-4 ketoreductase/SAM-dependent dTDP-4-keto-2,6-dideoxy-D-glucose C-3 C-methyltransferase [D-mycarose 3-C-methyltransferase]
MDIYGTARAVTTCPMCGAQDWQEVVDFGPVPLADSFLEPAASYDDEPRYPLAVVSCASCR
LMSLTHVVDPEVLYRTYPYTTSDSETIKKHMGHVVAGVRPSASASPEGSFVLEIGSNTGS
QLKAFQNAGMRTLGIDPARNIAAVANERGIETLPEFFSVDTAALVKKTHGTPQLVLGRHV
FAHIDDVSAVAEGVRDLLGPDSLFAIEVPYLVDMLERNEFDTIYHEHLSYIGVGSLVALF
RRHGLRVVDVERLAVHGGSILVFVGLDEGTRATAPVVEELIALEKERGLYEDATYERFAR
HVAEITAELTSMVRSLRAEGKRIAGYGAPAKGNTLLNVCGLTADDLEFCCDTTEFKQGLV
LPGTHIPVRSPEYAKTQAIDYYLLLAWNYGEEILAKEGPFLADGGRFILPNPRPSIVPPG
E
selected fasta
>NADPH-dependent dTDP-4-keto-2,6-dideoxy-D-glucose C-4 ketoreductase/SAM-dependent dTDP-4-keto-2,6-dideoxy-D-glucose C-3 C-methyltransferase [D-mycarose 3-C-methyltransferase]
ATGGACATCTATGGGACCGCGCGGGCAGTGACCACCTGCCCGATGTGCGGAGCGCAGGAC
TGGCAGGAGGTCGTGGACTTCGGCCCCGTCCCCCTGGCGGACAGCTTCCTCGAACCCGCC
GCGTCCTACGACGACGAACCGCGCTACCCGCTGGCCGTCGTCTCCTGCGCATCCTGCCGC
CTGATGAGCCTGACACACGTCGTGGACCCCGAAGTCCTGTACCGCACGTACCCCTACACC
ACCTCCGACTCGGAGACCATCAAGAAGCACATGGGGCATGTCGTCGCCGGTGTGCGTCCG
AGCGCTTCGGCATCCCCCGAGGGCAGCTTCGTCCTGGAGATCGGCAGCAACACCGGCTCC
CAGCTGAAGGCGTTCCAGAACGCCGGGATGCGCACCCTCGGCATCGACCCGGCCCGCAAC
ATCGCCGCGGTGGCCAACGAGCGGGGCATCGAGACGCTGCCGGAGTTCTTCTCGGTCGAC
ACCGCCGCCCTGGTGAAGAAGACCCACGGCACGCCCCAGCTGGTCCTCGGCCGGCATGTC
TTCGCGCACATCGACGACGTGTCCGCGGTCGCCGAGGGCGTCCGCGATCTGCTGGGCCCG
GACAGCCTGTTCGCCATCGAGGTGCCGTACCTGGTGGACATGCTGGAGCGCAACGAGTTC
GACACCATCTACCACGAGCACCTGTCCTACATCGGCGTCGGCTCCCTGGTCGCCCTCTTC
CGACGGCACGGGCTGCGTGTGGTGGACGTCGAACGCCTCGCGGTGCACGGCGGATCCATC
CTGGTCTTCGTCGGGCTGGACGAGGGCACCCGGGCGACGGCGCCCGTCGTGGAGGAGCTG
ATCGCGCTGGAGAAGGAGCGCGGTCTGTACGAGGACGCCACCTACGAGCGGTTCGCCAGG
CACGTCGCGGAGATCACCGCCGAACTGACCTCGATGGTCCGCTCGCTGCGGGCCGAGGGC
AAACGGATCGCCGGGTACGGGGCGCCCGCGAAGGGCAACACCCTGCTGAACGTCTGCGGG
CTCACCGCGGACGACCTGGAATTCTGCTGCGACACCACCGAGTTCAAGCAGGGCCTGGTG
CTGCCCGGCACGCACATCCCGGTCCGCAGCCCCGAGTACGCCAAGACCCAGGCCATCGAC
TACTACCTGCTGCTGGCCTGGAACTACGGCGAGGAGATCCTCGCCAAGGAGGGGCCGTTC
CTGGCGGACGGCGGACGGTTCATCCTGCCCAACCCGCGGCCGTCGATCGTCCCGCCCGGG
GAGTGA

close this sectionFeature

BLASTP
Database:UniProtKB:2011_09 		show BLAST table
InterPro
Database:interpro:38.0
IPR013630 Methyltransferase putative zinc binding domain (Domain)
[13-75] 7e-21 PF08421
PF08421   Methyltransf_13
IPR013691 C-methyltransferase (Domain)
[254-411] 9e-55 PF08484
PF08484   Methyltransf_14
SignalP No significant hit
TMHMM No significant hit
Mith_00200 Mith_00200   Mith_00220 Mith_00220
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