Product chain-length determination mechanism of Z,E-farnesyl diphosphate synthase.
cis-Prenyltransferases catalyze the consecutive condensation of isopentenyl diphosphate (IPP) with allylic prenyl diphosphates, producing Z,E-mixed prenyl diphosphate. The Mycobacterium tuberculosis Z,E-farnesyl diphosphate synthase Rv1086 catalyzes the condensation of one molecule of IPP with geranyl diphosphate to yield Z,E-farnesyl diphosphate and is classified as a short-chain cis-prenyltransferase. To elucidate the chain-length determination mechanism of the short-chain cis-prenyltransferase, we introduced some substitutive mutations at the characteristic amino acid residues of Rv1086. Among the mutants constructed, L84A showed a dramatic change of catalytic function to synthesize longer prenyl chain products than that of wild type, indicating that Leu84 of Rv1086 plays an important role in product chain-length determination. Mutagenesis at the corresponding residue of a medium-chain cis-prenyltransferase, Micrococcus luteus B-P 26 undecaprenyl diphosphate synthase also resulted in the production of different prenyl chain length from the intrinsic product, suggesting that this position also plays an important role in product chain-length determination for medium-chain cis-prenyltransferases.
Noike
Motoyoshi
M
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai, Miyagi, Japan.
Ambo
Takanori
T
Kikuchi
Sayaka
S
Suzuki
Toshihide
T
Yamashita
Satoshi
S
Takahashi
Seiji
S
Kurokawa
Hirofumi
H
Mahapatra
Sebabrata
S
Crick
Dean C
DC
Koyama
Tanetoshi
T
D019883:Alkyl and Aryl Transferases / Q000737:chemistry* / Q000235:genetics; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D002384:Catalysis; D017124:Conserved Sequence; D004122:Dimethylallyltranstransferase / Q000737:chemistry / Q000235:genetics; D051228:Geranyltranstransferase / Q000737:chemistry* / Q000235:genetics; D007930:Leucine / Q000737:chemistry* / Q000235:genetics; D016982:Micrococcus luteus / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D017433:Protein Structure, Secondary / Q000235:genetics; D016415:Sequence Alignment
D001426:Bacterial Proteins; D019883:Alkyl and Aryl Transferases; C412569:Rv1086 protein, Mycobacterium tuberculosis; D004122:Dimethylallyltranstransferase; D051228:Geranyltranstransferase; D007930:Leucine
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