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PMID: 10536025 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Interactions among enzymes of the Arabidopsis flavonoid biosynthetic pathway.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 22 ·1999-10-26 ·Pages 12929-34

Burbulis IE, Winkel-Shirley B

Abstract

Flavonoids are secondary metabolites derived from phenylalanine and acetate metabolism that perform a variety of essential functions in higher plants. Studies over the past 30 years have supported a model in which flavonoid metabolism is catalyzed by an enzyme complex localized to the endoplasmic reticulum [Hrazdina, G. & Wagner, G. J. (1985) Arch. Biochem. Biophys. 237, 88-100]. To test this model further we assayed for direct interactions between several key flavonoid biosynthetic enzymes in developing Arabidopsis seedlings. Two-hybrid assays indicated that chalcone synthase, chalcone isomerase (CHI), and dihydroflavonol 4-reductase interact in an orientation-dependent manner. Affinity chromatography and immunoprecipitation assays further demonstrated interactions between chalcone synthase, CHI, and flavonol 3-hydroxylase in lysates from Arabidopsis seedlings. These results support the hypothesis that the flavonoid enzymes assemble as a macromolecular complex with contacts between multiple proteins. Evidence was also found for posttranslational modification of CHI. The importance of understanding the subcellular organization of elaborate enzyme systems is discussed in the context of metabolic engineering.

MeSH Terms
Arabidopsis/enzymology,metabolism Base Sequence Chromatography, Affinity DNA Primers Enzymes/isolation & purification,metabolism Flavonoids/biosynthesis Precipitin Tests Protein Binding
Chemicals
DNA Primers Enzymes Flavonoids
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Burbulis I E
Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061-0406, USA.
Winkel-Shirley B
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.22.12929
Published
1999-10-26
Pages
12929-34
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC23169
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.22.12929 PMC Full Text View journal details
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