Tissue and method specificities of phenylalanine ammonia-lyase assay
Language English Country Germany Media print-electronic
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
22677451
DOI
10.1016/j.jplph.2012.04.008
PII: S0176-1617(12)00193-9
Knihovny.cz E-resources
- MeSH
- Arabidopsis enzymology MeSH
- Phenols metabolism MeSH
- Phenylalanine Ammonia-Lyase metabolism MeSH
- Plant Roots enzymology MeSH
- Plant Leaves enzymology MeSH
- Matricaria enzymology MeSH
- Copper metabolism MeSH
- Spectrophotometry * MeSH
- Substrate Specificity MeSH
- Chromatography, High Pressure Liquid * MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Names of Substances
- Phenols MeSH
- Phenylalanine Ammonia-Lyase MeSH
- Copper MeSH
A large number of studies have estimated phenylalanine ammonia-lyase (PAL) activity because it strongly reacts to various stimuli. Activity of this enzyme has been assayed mainly by means of spectrophotometry, but the precision of this method is poorly known. We compared assays of PAL activity using spectrophotometry and high performance liquid chromatography (HPLC) in two species (Matricaria chamomilla and Arabidopsis thaliana). Additionally, copper-exposed M. chamomilla plants and buffer with additive were also tested. Our data indicate that spectrophotometry both overestimates (leaves of M. chamomilla) and underestimates (leaves and roots of A. thaliana) PAL activity in comparison with HPLC, suggesting interference of UV-absorbing metabolites. HPLC also showed more accurate detection of cinnamic acid in Cu-exposed chamomile roots. Addition of dithiothreitol to the extraction buffer enhanced PAL activity but reduced proteins, indicating an artificial negative effect. A comparison of PAL activity in selected species is also provided.
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