, Volume 169, Issue 1, pp 97–107 | Cite as

L-Phenylalanine ammonia-lyase fromPhaseolus vulgaris: Modulation of the levels of active enzyme bytrans-cinnamic acid

  • G. P. Bolwell
  • C. L. Cramer
  • C. J. Lamb
  • W. Schuch
  • R. A. Dixon


The extractable activity ofl-phenylalanine ammonia-lyase (PAL; EC in cell suspension cultures of bean (Phaseolus vulgaris) is greatly induced following exposure to an elicitor preparation from the cell walls of the phytopathogenic fungusColletotrichum lindemuthianum. Following exogenous application oftrans-cinnamic acid (the product of the PAL reaction) to elicitor-induced cells, the activity of the enzyme rapidly declines. Loss of enzyme activity is accompanied by inhibition of the rate of synthesis of PAL subunits, as determined by [35S]methionine pulse-labelling followed by specific immunoprecipitation; this is insufficient to account for the rapid loss of PAL enzyme activity. Pulse-chase and immune blotting experiments indicate that cinnamic acid does not affect the rate of degradation of enzyme subunits, but rather mediates inactivation of the enzyme. A non-dialysable factor from cinnamicacid-treated bean cells stimulates removal of PAL activity from enzyme extracts in vitro; this effect is dependent on the presence of cinnamic acid. Such loss of enzyme activity in vitro is accompanied by an apparent loss or reduction of the dehydroalanine residue of the enzyme's active site, as detected by active-site-specific tritiation, although levels of immunoprecipitable enzyme subunits do not decrease. Furthermore, cinnamic-acid-mediated loss of enzyme activity in vivo is accompanied, in pulse-chase experiments, by a greater relative loss of35S-labelled enzyme subunits precipitated by an immobilised active-site affinity ligand than of subunits precipitated with anti-immunoglobulin G. It is therefore suggested that a possible mechanism for cinnamic-acid-mediated removal of PAL activity may involve modification of the dehydroalanine residue of the enzyme's active site.

Key words

Cell culture Phaseolus (enzyme turnover) Phenylalanine ammonia-lyase Phenylpropanoid biosynthesis 



l-α-aminoxy-β-phenylpropionic acid


trans-cinnamic acid


polyacrylamide gel electrophoresis


l-phenylalanine ammonia-lyase


sodium dodecyl sulphate


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Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • G. P. Bolwell
    • 1
  • C. L. Cramer
    • 2
  • C. J. Lamb
    • 2
  • W. Schuch
    • 3
  • R. A. Dixon
    • 1
  1. 1.Department of Biochemistry, Royal Holloway and Bedford New CollegeUniversity of LondonEghamUK
  2. 2.Plant Biology LaboratorySalk Institute for Biological StudiesSan DiegoUSA
  3. 3.Imperial Chemical Industries plc, Corporate Biosciences LaboratoryThe HealthRuncornUK

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