Purification, kinetics, inhibitors and CD for recombinant β-amyrin synthase from Euphorbia tirucalli L and functional analysis of the DCTA motif, which is highly conserved among oxidosqualene cyclases
Article first published online: 13 FEB 2013
© 2013 The Authors Journal compilation © 2013 FEBS
Volume 280, Issue 5, pages 1267–1280, March 2013
How to Cite
Ito, R., Masukawa, Y. and Hoshino, T. (2013), Purification, kinetics, inhibitors and CD for recombinant β-amyrin synthase from Euphorbia tirucalli L and functional analysis of the DCTA motif, which is highly conserved among oxidosqualene cyclases. FEBS Journal, 280: 1267–1280. doi: 10.1111/febs.12119
- Issue published online: 26 FEB 2013
- Article first published online: 13 FEB 2013
- Accepted manuscript online: 7 JAN 2013 12:50PM EST
- Manuscript Accepted: 1 JAN 2013
- Manuscript Revised: 23 DEC 2012
- Manuscript Received: 8 NOV 2012
- Ministry of Education, Culture, Sports, Science and Technology, Japan. Grant Number: 18380001
Fig. S1. Structure determination of product 3 acetate by the detailed NMR analyses.
Fig. S2. Structure determination of product 4 acetate by the detailed NMR analyses.
Fig. S3. Determination of molecular mass by SDS/PAGE.
Fig. S4. (A) CD spectra measured in the solutions of Brij 35 and beta-OG and (B) the enzyme activity recovered after removing Brij35.
Fig. S5. Effect of dithiothreitol on enzymatic activity.
Fig. S6. Synthetic scheme of (3R,S)-iminosqualene (5).
Fig. S7. Determination of IC50 values of inhibitors 5–7.
Fig. S8. Amino acid alignment of various oxidosqualene cyclases.
Fig. S9. The method for determining the expressed amounts of EtAS proteins.
Table S1. Active site residues of human lanosterol synthase that interact with the inhibitor Ro48-8071 (6).
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