The role of histidine‐114 of Sulfolobus acidocaldarius geranylgeranyl diphosphate synthase in chain‐length determination

FEBS Letters

Kato, Tatsuya¹  Hemmi, Hisashi¹  Hirooka, Kazutake¹  Nishino, Tokuzo¹  Matsu-ura, Jun-ichiro¹ 

[1] Department of Biochemistry and Engineering, Tohoku University, Aoba Aramaki 07, Aoba-ku, Sendai 980-8579, Japan

关键词: Prenyltransferase;    Geranylgeranyl diphosphate;    Product specificity;    Site-directed mutagenesis;    Molecular modeling;    GGPP;    (all-E)-geranylgeranyl diphosphate;    GGPS;    geranylgeranyl diphosphate synthase;    IPP;    isopentenyl diphosphate;    DMAPP;    dimethylallyl diphosphate;    GPP;    geranyl diphosphate;    FPP;    (all-E)-farnesyl diphosphate;    FARM;    first aspartate-rich motif;    TLC;    thin-layer chromatography;   

DOI  :  10.1016/S0014-5793(00)01972-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Sulfolobus acidocaldarius geranylgeranyl diphosphate synthase yields (all-E)-C₂₀ prenyl diphosphate as a final product. The three-dimensional model of the enzyme suggested that removing two bulky residues at 77 and 114 would allow additional prenyl-chain elongation. To test this, we examined several mutants with substitutions at 77 and/or 114. As a result, the mutants, F77G, F77G and H114A, F77G and H114G, H114A, and H114G gave C₃₀, C₄₅, C₅₀, C₃₀ and C₄₀ as the main long product, respectively. These observations indicate that histidine-114 plays a crucial role in chain-length determination along with phenylalanine-77.

【 授权许可】

Unknown