Ebony, a Novel Nonribosomal Peptide Synthetase for β-Alanine Conjugation with Biogenic Amines in Drosophila

Using Ebony protein either expressed in Escherichia coli or in Schneider S2 cells, we provide evidence for its substrate specificity and reaction mechanism. Ebony activates β-alanine to aminoacyladenylate by an adenylation domain and covalently attaches it as a thioester to a thiolation domain in a...

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Published in:The Journal of biological chemistry 2003-10, Vol.278 (42), p.41160
Main Authors: Arnd Richardt, Tobias Kemme, Stefanie Wagner, Dirk Schwarzer, Mohamed A. Marahiel, Bernhard T. Hovemann
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container_issue 42
container_start_page 41160
container_title The Journal of biological chemistry
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creator Arnd Richardt
Tobias Kemme
Stefanie Wagner
Dirk Schwarzer
Mohamed A. Marahiel
Bernhard T. Hovemann
description Using Ebony protein either expressed in Escherichia coli or in Schneider S2 cells, we provide evidence for its substrate specificity and reaction mechanism. Ebony activates β-alanine to aminoacyladenylate by an adenylation domain and covalently attaches it as a thioester to a thiolation domain in a n on r ibosomal p eptide s ynthetase (NRPS) related mechanism. In a second reaction, biogenic amines act as external nucleophiles on β-alanyl- S -pantetheine-Ebony, thereby releasing in a fast reaction the dipeptide (peptidoamine) in a process that is novel in higher eucaryotes. Therefore, we define Ebony as a β-alanyl-biogenic amine synthetase. Insight into the reaction mechanism stems from mutational analysis of an invariant serine that disclosed Ebony as a multienzyme with functional analogy to the starting modules of NRPSs. In light of a putative biogenic amine-deactivating capacity, Ebony function in the nervous system must be reconsidered. We propose that in the Drosophila eye Ebony is involved in the transmission process by inactivation of histamine through β-alanyl conjugation.
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title Ebony, a Novel Nonribosomal Peptide Synthetase for β-Alanine Conjugation with Biogenic Amines in Drosophila
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