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Alkaloid N-oxides as transport and vacuolar storage compounds of pyrrolizidine alkaloids in Senecio vulgaris L

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Abstract

Cell-suspension cultures of pyrrolizidinealkaloid-producing species selectively take up and accumulate senecionine (sen) and its N-oxide (sen-Nox). Cultures established from non-alkaloid-producing species are unable to accumulate the alkaloids. The uptake and accumulation of 14C-labelled alkaloids was studied using a Senecio vulgaris cell-suspension culture as well as protoplasts and vacuoles derived from it. The alkaloid uptake exhibits all characteristics of a carrier-mediated transport. The uptake of sen-Nox follows a multiphasic saturation kinetics. The Km-values for sen Nox of 53 μM and 310 μM are evaluated. Senecionine competitively inhibits sen-Nox uptake, indicating that the tertiary alkaloid and its N-oxide share the same membrane carrier. The N-oxide of sen shows a pH optimum below 5.5, whereas sen is taken up over a range from pH 4 to 8. Activation energies of 90 and 53 kJ·mol-1 are calculated for sen-Nox and sen transport, respectively. At concentrations of 10 to 100 μM, sen-Nox is rapidly taken up by cells and protoplasts; within 2 h >90% of total N-oxide is within the cells. By contrast the uptake of sen is less efficient. Vacuoles isolated from protoplasts preloaded with sen-Nox totally retained the alkaloid N-oxide, whereas sen is rapidly lost during the procedure of vacuole preparation. N-oxidation converts the weak lipophilic tertiary base into a charged polar molecule which is excellently adapted to serve as the cellular transport and storage form of pyrrolizidine alkaloids.

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Abbreviations

CCCP:

carbonylcyanide m-chlorophenylhydrazone

DCCD:

N,N′-dicyclohexylcarbodiimide

DIDS:

4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid

DNP:

2,4-dinitrophenol

sen:

senecionine

sen-Nox:

senecionine N-oxide

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  1. Institut für Pharmazeutische Biologie der Technischen Universität Braunschweig, Mendelssohnstrasse 1, D-3300, Braunschweig, Federal Republic of Germany

    Adelheid Ehmke, Kirsten von Borstel & Thomas Hartmann

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  1. Adelheid Ehmke
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  2. Kirsten von Borstel
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  3. Thomas Hartmann
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Ehmke, A., von Borstel, K. & Hartmann, T. Alkaloid N-oxides as transport and vacuolar storage compounds of pyrrolizidine alkaloids in Senecio vulgaris L. Planta 176, 83–90 (1988). https://doi.org/10.1007/BF00392483

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