A project at 
Project
summary: Plants biosynthesize an amazing array
of structurally diverse small molecules. The purpose
of this multidisciplinary collaborative research is to determine
how and why plants synthesize diverse triterpenoids.
This project is integrating reverse genetics in Arabidopsis,
heterologous expression of cDNAs in yeast, spectroscopic
and chromatographic structural determination, and gene expression
analysis to elucidate the function of Arabidopsis triterpenoid
biosynthetic genes and thereby triterpenoid biosynthetic
pathways, control mechanisms, and biological function.
Three of the characterized triterpenoid biosynthetic enzymes
convert a shared substrate to different compounds; other
enzymes to be studied may similarly provide structural diversity,
may metabolize different substrates, or may provide means
to spatially or temporally control expression and thus product
formation. The effects of the mutations on plant growth,
development, and triterpenoid composition will allow linkage
of each triterpenoid biosynthetic gene with an enzyme of
known catalytic activity and a specific biological role.
Enzymatic
synthesis of an indole diterpene by an oxidosqualene cyclase:
Mechanistic, biosynthetic, and phylogenetic implications.
Xiong, Q., Zhu, X., Wilson, W.K., Ganesan,
A., and Matsuda, S.P.T. J. Am. Chem., 125 (2003): 9002-9003.
Abstract;
PDF
Mutagenesis
approaches to deduce structure-function relationships in terpene
synthases.
M. J. R. Segura, B. E. Jackson, and S. P. T. Matsuda. Nat.
Prod. Rep., 20 (2003): 304-317.
Abstract
Subcellular
localization of oxidosqualene cyclases from Arabidopsis thaliana,
Trypanosoma cruzi, and Pneumocystis carinii expressed
in yeast.
P. Milla, F. Viola, S. Oliaro-Bosso, F.
Rocco, L. Cattel, B. M. Joubert, R. J. LeClair, Matsuda, S. P.
T., and G. Balliano. Lipids, 37 (2002): 1171-1176.
Abstract
Directed
evolution experiments reveal mutations at cycloartenol synthase
residue His477 that dramatically alter catalysis.
M. J. R. Segura, S. Lodeiro, M. M. Meyer, A. J. Patel, and S.
P. T. Matsuda. Org. Lett., 4 (2002): 4459-4462.
Abstract;
PDF
Directed
evolution to generate cycloartenol synthase mutants that produce
lanosterol.
M. M. Meyer, R. Xu, and S. P. T. Matsuda. Org. Lett., 4
(2002):1395-1398.
Abstract;
PDF
Functional
cloning of an Arabidopsis thaliana a cDNA encoding cycloeucalenol
cycloisomerase.
M. A. Lovato, E. A. Hart, M. J. R. Segura, J.-L. Giner, and S.
P. T. Matsuda. J. Biol. Chem., 275 (2000): 13394-13397.
Abstract;
PDF
A
tyrosine to threonine mutation converts cycloartenol synthase
to an oxidosqualene cyclase that forms lanosterol as its major
product.
J. B. R. Herrera, W. K. Wilson, and S. P. T. Matsuda. J.
Am. Chem. Soc., 122 (2000): 6765-6766.
PDF
Arabidopsis
thaliana LUP1 converts oxidosqualene to multiple triterpene
alcohols and a triterpene diol.
M. J. R. Segura, M. M. Meyer, and S. P. T. Matsuda. Org.
Lett., 2 (2000): 2257-2259.
Abstract;
PDF
Steric
bulk at cycloartenol synthase position 481 influences cyclization
and deprotonation.
S. P. T. Matsuda, L. B. Darr, E. A. Hart, J. B. R. Herrera, K.
E. McCann, M. M. Meyer, J. Pang, H. G. Schepmann, and W. K. Wilson. Org. Lett., 2 (2000): 2261-2263.
Abstract;
PDF
Oxidosqualene
cyclase residues that promote formation of cycloartenol, lanosterol,
and parkeol.
M.M. Meyer, M.J.R. Segura, W.K.Wilson, and S.P.T. Matsuda. Angew. Chem., 39 (2000): 4090-4092.
PDF
Directed
evolution to investigate steric control of enzymatic oxidosqualene
cyclization. An isoleucine to valine mutation in cycloartenol
synthase allows lanosterol and parkeol biosynthesis.
E. A. Hart, L. Hua, L. B. Darr, W. K. Wilson, J. Pang, and S.
P. T. Matsuda. J. Am. Chem. Soc., 121 (1999): 9887-9888.
PDF
Cloning
and characterization of the Arabidopsis thaliana lupeol
synthase gene.
Herrera, J.B.R., Bartel, B., Wilson, W.K., and Matsuda, S.P.T.
(1998) Phytochemistry 49, 1905-1911.
Abstract;full
text; PDF
Isolation
of an Arabidopsis thaliana gene encoding cycloartenol synthase
by functional expression in a yeast mutant lacking lanosterol
synthase by the use of a chromatographic screen.
Corey, E.J., Matsuda, S.P.T., and Bartel, B. (1993) Proc. Natl.
Acad. Sci. USA 90, 11628-11632.
Abstract;
PDF
