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Plant and Cell Physiology, 1990, Vol. 31, No. 2 223-231
© 1990

Article

Folylpolyglutamate Derivatives of Pisum sativum L. Determination of Polyglutamate Chain Lengths by High Performance Liquid Chromatography Following Conversion to p-aminobenzoylpolyglutamates

Helena C. Imeson1, Li-li Zheng2 and Edwin A. Cossins3

Department of Botany, University of Alberta Edmonton, Alberta, T6G 2E9, Canada

3To whom correspondence should be addressed

The folypolyglutamate derivatives of pea seedlings (Pisum sativum L. cv. Homesteader) were extracted in the presence of 2-mercaptoethanol and cleaved to p-aminobenzoylpolyglutamates by treatment with Zn-HCl. Azo dyes were formed by reaction with naphthylethylene diamine and purified by polyacrylamide gel chromatography. p-Aminobenzoylpolyglutamates were regenerated from these dyes by Zn treatment and then concentrated in vacuo. These derivatives were separated according to glutamyl chain length by high performance liquid chromatography on Whatman Partisil SAX columns. The folylpolyglutamates of 4 day old pea cotyledons, pea leaves and isolated chloroplasts were mainly tetra- and pentaglutamates. These and folates of shorter chain length were labelled when seeds and aerial shoots were incubated with p-aminobenzoate-[14C]. Labelling of the pentaglutamate was reduced in seeds that were imbibed in the presence of 0.1 mM methotrexate. Studies of cotyledon folylpolyglutamate synthetase showed that polyglutamate chain length was affected by incubation time and the concentration of tetra-hydrofolate monoglutamate in the reaction system.

1Present address: Department of Biology, University of Lethbridge, Alberta, Canada T1K 3M4

2Present address: Department of Horticulture, Xiong-yue Agricultural College, Xiong-yue, Liaoning Province, China

(Received August 4, 1989; Accepted December 5, 1989)
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