6WW1

Crystal structure of the LmFPPS mutant E97Y

6WW1 の概要

• エントリーDOI: 10.2210/pdb6ww1/pdb
• 分子名称: Farnesyl pyrophosphate synthase, CALCIUM ION, ISOPENTYL PYROPHOSPHATE, ... (6 entities in total)
• 機能のキーワード: fpps, farnesyl diphosphate synthase, transferase
• 由来する生物種: Leishmania major
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 83605.73

構造登録者

Maheshwari, S.,Kim, Y.S.,Gabelli, S.B. (登録日: 2020-05-07, 公開日: 2020-10-07, 最終更新日: 2023-10-18)

主引用文献

Maheshwari, S.,Kim, Y.S.,Aripirala, S.,Murphy, M.,Amzel, L.M.,Gabelli, S.B.
Identifying Structural Determinants of Product Specificity in Leishmania major Farnesyl Diphosphate Synthase.
Biochemistry, 59:2751-2759, 2020

PubMed Abstract: Farnesyl diphosphate synthase (FPPS) is an isoprenoid chain elongation enzyme that catalyzes the sequential condensation of dimethylallyl diphosphate (C) with isopentenyl diphosphate (IPP; C) and the resulting geranyl diphosphate (GPP; C) with another molecule of IPP, eventually producing farnesyl diphosphate (FPP; C), which is a precursor for the biosynthesis of a vast majority of isoprenoids. Previous studies of FPPS have highlighted the importance of the structure around the hydrophobic chain elongation path in determining product specificity. To investigate what structural features define the final chain length of the product in FPPS from , we designed and expressed six mutants of FPPS by replacing small amino acids around the binding pocket with bulky residues. Using enzymatic assays, binding kinetics, and crystallographic studies, we analyzed the effects of these mutations on the activity and product specificity of FPPS. Our results revealed that replacement of Thr-164 with tryptophan and phenylalanine completely abolished the activity of FPPS. Intriguingly, the T164Y substitution displayed dual product specificity and produced a mixture GPP and FPP as final products, with an activity for FPP synthesis that was lower than that of the wild-type enzyme. These data indicate that Thr-164 is a potential regulator of product specificity.

PubMed: 32584028
DOI: 10.1021/acs.biochem.0c00432

実験手法

X-RAY DIFFRACTION (2.05 Å)