TY - JOUR
T1 - Product chain-length determination mechanism of Z,E-farnesyl diphosphate synthase
AU - Noike, Motoyoshi
AU - Ambo, Takanori
AU - Kikuchi, Sayaka
AU - Suzuki, Toshihide
AU - Yamashita, Satoshi
AU - Takahashi, Seiji
AU - Kurokawa, Hirofumi
AU - Mahapatra, Sebabrata
AU - Crick, Dean C.
AU - Koyama, Tanetoshi
N1 - Funding Information:
This work was supported in part by grants-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant 19710178 to M.N.) and from Tohoku University Exploratory Research Program for Young Scientists (TU-ERYS) and from NIH/NIAID Grants AI-065357 and AI-018357.
PY - 2008/12/5
Y1 - 2008/12/5
N2 - 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.
AB - 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.
KW - Chain-length determination mechanism
KW - Isoprenoid
KW - Mutagenesis
KW - Mycobacterium tuberculosis
KW - Prenyl diphosphate synthase
KW - Prenyltransferase
KW - Undecaprenyl diphosphate synthase
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U2 - 10.1016/j.bbrc.2008.09.014
DO - 10.1016/j.bbrc.2008.09.014
M3 - Article
C2 - 18790692
AN - SCOPUS:54449097408
SN - 0006-291X
VL - 377
SP - 17
EP - 22
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -
