In:
ChemBioChem, Wiley, Vol. 18, No. 21 ( 2017-11-02), p. 2145-2155
Kurzfassung:
The functions of Val483, Trp534, and Met729 in Euphorbia tirucalli β‐amyrin synthase were revealed by comparing the enzyme activities of site‐directed mutants against that of the wild type. The Gly and Ala variants with a smaller bulk size at position 483 predominantly afforded monocyclic camelliol C, which suggested that the orientation of the (3 S )‐2,3‐oxidosqualene substrate was not appropriately arranged in the reaction cavity as a result of the decreased bulk size, leading to failure of its normal folding into the chair–chair–chair–boat–boat conformation. The Ile variant, with a somewhat larger bulk, afforded β‐amyrin as the dominant product. Intriguingly, various variants of Trp534 exhibited significantly decreased enzymatic activities and provided no aberrantly cyclized products, although the aromatic Phe and Tyr residues were incorporated and the steric sizes of the aliphatic residues were altered. Therefore, the Trp534 residue does not stabilize the transient cation through a cation–π interaction. Furthermore, the Trp residue, with the largest steric bulk among all natural amino acids, is essential for high enzymatic activity. Robust CH–π complexation between the Val483 and Trp534 residues is proposed herein. Altering the steric bulk at the Met729 position afforded the pentacyclic skeletons. Thus, Met729 is positioned at the E‐ring formation site. More detailed insights into the functions of the Val483, Trp534, and Met729 residues are provided by homology modeling.
Materialart:
Online-Ressource
ISSN:
1439-4227
,
1439-7633
DOI:
10.1002/cbic.201700368
Sprache:
Englisch
Verlag:
Wiley
Publikationsdatum:
2017
ZDB Id:
2020469-3
SSG:
12
