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J Biol Chem, Vol. 274, Issue 7, 4273-4280, February 12, 1999
From the Hydroxycinnamoyl-CoA:tyramine
N-(hydroxycinnamoyl)transferase (THT; EC 2.3.1.110)
catalyzes the transfer of hydroxycinnamic acids from the respective CoA
esters to tyramine and other amines in the formation of
N-(hydroxycinnamoyl)amines. Expression of THT is induced by
Phytophthora infestans, the causative agent of late blight
disease in potato. The amino acid sequences of nine endopeptidase
LysC-liberated peptides from purified potato THT were determined. Using
degenerate primers, a THT-specific fragment was obtained by reverse
transcription-polymerase chain reaction, and THT cDNA clones were
isolated from a library constructed from RNA of elicitor-treated potato
cells. The open reading frame encoding a protein of 248 amino acids was
expressed in Escherichia coli. Recombinant THT exhibited a
broad substrate specificity, similar to that of native potato THT,
accepting cinnamoyl-, 4-coumaroyl-, caffeoyl-, feruloyl- and
sinapoyl-CoA as acyl donors and tyramine, octopamine, and noradrenalin
as acceptors tested. Elicitor-induced THT transcript accumulation in
cultured potato cells peaked 5 h after initiation of treatment,
whereas enzyme activity was highest from 5 to 30 h after
elicitation. In soil-grown potato plants, THT mRNA was most
abundant in roots. Genomic Southern analyses indicate that, in potato,
THT is encoded by a multigene family.
Cloning and Expression of a Potato cDNA Encoding
Hydroxycinnamoyl-CoA:Tyramine
N-(Hydroxycinnamoyl)transferase
,
,, and
Abteilung Sekundärstoffwechsel,
¶ Abteilung Naturstoffchemie and Abteilung Stress- und
Entwicklungsbiologie of the Institut für Pflanzenbiochemie (IPB),
Weinberg 3, D-06120 Halle (Saale), Germany, and § Waldbronn
Analytical Division, Hewlett-Packard GmbH, Hewlett-Packard Str. 8, D-76337 Waldbronn, Germany
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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