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Characterization of trimeric acetylcholinesterase from a legume plant, Macroptilium atropurpureum Urb.

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Abstract

We recently identified plant acetylcholinesterases (E.C.3.1.1.7; AChEs) homologous to the AChE purified from a monocotyledon, maize, that are distinct from the animal AChE family. In this study, we purified, cloned and characterized an AChE from a dicotyledon, siratro. The full-length cDNA of siratro AChE is 1,441 nucleotides, encoding a 382-residue protein that includes a signal peptide. This AChE is a disulfide-linked 125-kDa homotrimer consisting of 41–42 kDa subunits, in contrast to the maize AChE, which exists as a mixture of disulfide and non-covalently linked 88-kDa homodimers. The plant AChEs apparently consist of various quaternary structures, depending on the plant species, similar to the animal AChEs. We compared the enzymatic properties of the dimeric maize and trimeric siratro AChEs. Similar to electric eel AChE, both plant AChEs hydrolyzed acetylthiocholine (or acetylcholine) and propionylthiocholine (or propionylcholine), but not butyrylthiocholine (or butyrylcholine), and their specificity constant was highest against acetylcholine. There was no significant difference between the enzymatic properties of trimeric and dimeric AChEs, although two plant AChEs had low substrate turnover numbers compared with electric eel AChE. The two plant AChE activities were not inhibited by excess substrate concentrations. Thus, similar to some plant AChEs, siratro and maize AChEs showed enzymatic properties of both animal AChE and animal BChE. On the other hand, both siratro and maize AChEs exhibited low sensitivity to the AChE-specific inhibitor neostigmine bromide, dissimilar to other plant AChEs. These differences in enzymatic properties of plant AChEs may reflect the phylogenetic evolution of AChEs.

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Abbreviations

ACh:

Acetylcholine

PpCh:

Propionylcholine

BCh:

Butyrylcholine

ASCh:

Acetylthiocholine

PpSCh:

Propionylthiocholine

BSCh:

Butyrylthiocholine

AChE:

Acetylcholinesterase

DTNB:

5, 5′-Dithiobis-2-nitrobenzoic acid

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Acknowledgments

This work was supported by Japan Space Forum. Sequence data from this article have been deposited with the DDBJ/EMBL/GenBank data libraries under accession number AB294246.

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Authors and Affiliations

  1. Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, Hokkaido, 099-2493, Japan

    Kosuke Yamamoto, Suguru Oguri & Yoshie S. Momonoki

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  1. Kosuke Yamamoto
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  2. Suguru Oguri
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  3. Yoshie S. Momonoki
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Correspondence to Yoshie S. Momonoki.

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Kosuke Yamamoto and Yoshie S. Momonoki contributed equally to this work.

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Yamamoto, K., Oguri, S. & Momonoki, Y.S. Characterization of trimeric acetylcholinesterase from a legume plant, Macroptilium atropurpureum Urb.. Planta 227, 809–822 (2008). https://doi.org/10.1007/s00425-007-0658-0

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