Biomaterial Database

Tissue and method specificities of phenylalanine ammonia-lyase assay. 2012

Jozef Kováčik, and Bořivoj Klejdus

Institute of Chemistry and Biochemistry, Mendel University in Brno, Zemědělská 1, 613 00 Brno, Czech Republic. jozkovacik@yahoo.com

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.

UI	MeSH Term	Description	Entries
D010636	Phenols	Benzene derivatives that include one or more hydroxyl groups attached to the ring structure.
D010650	Phenylalanine Ammonia-Lyase	An enzyme that catalyzes the deamination of PHENYLALANINE to form trans-cinnamate and ammonia.	Phenylalanine-Tyrosine Ammonia-lyase,Ammonia-Lyase, Phenylalanine,Ammonia-lyase, Phenylalanine-Tyrosine,Phenylalanine Ammonia Lyase,Phenylalanine Tyrosine Ammonia lyase
D002851	Chromatography, High Pressure Liquid	Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.	Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D003300	Copper	A heavy metal trace element with the atomic symbol Cu, atomic number 29, and atomic weight 63.55.	Copper-63,Copper 63
D013053	Spectrophotometry	The art or process of comparing photometrically the relative intensities of the light in different parts of the spectrum.
D013379	Substrate Specificity	A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.	Specificities, Substrate,Specificity, Substrate,Substrate Specificities
D017360	Arabidopsis	A plant genus of the family BRASSICACEAE that contains ARABIDOPSIS PROTEINS and MADS DOMAIN PROTEINS. The species A. thaliana is used for experiments in classical plant genetics as well as molecular genetic studies in plant physiology, biochemistry, and development.	Arabidopsis thaliana,Cress, Mouse-ear,A. thaliana,A. thalianas,Arabidopses,Arabidopsis thalianas,Cress, Mouse ear,Cresses, Mouse-ear,Mouse-ear Cress,Mouse-ear Cresses,thaliana, A.,thaliana, Arabidopsis,thalianas, A.
D018515	Plant Leaves	Expanded structures, usually green, of vascular plants, characteristically consisting of a bladelike expansion attached to a stem, and functioning as the principal organ of photosynthesis and transpiration. (American Heritage Dictionary, 2d ed)	Plant Leaf,Leaf, Plant,Leave, Plant,Leaves, Plant,Plant Leave
D018517	Plant Roots	The usually underground portions of a plant that serve as support, store food, and through which water and mineral nutrients enter the plant. (From American Heritage Dictionary, 1982; Concise Dictionary of Biology, 1990)	Plant Bulbs,Plant Root,Bulb, Plant,Bulbs, Plant,Plant Bulb,Root, Plant,Roots, Plant
D031168	Matricaria	A plant genus of the family ASTERACEAE. M. chamomilla appears similar to Anthemis but this flower disk is conical and hollow and lacks chaffy bract scales and the odor is weaker. The common name of 'manzanilla' is confused with other meanings of the word. 'Matricaria chamomilla sensu' is classified by some as Tripleurospermum perforata. Other plants with similar common names include CHAMAEMELUM; TRIPLEUROSPERMUM and ANTHEMIS.	Camomile, German,Chamomile, German,Chamomilla recutita,Matricaria chamomilla,Matricaria recutita,Mayweed, Crown,Pineapple Weed,German Chamomile,Matricaria matricarioides,Chamomilla recutitas,Crown Mayweed,Crown Mayweeds,German Camomile,Matricaria chamomillas,Matricaria matricarioide,Matricaria recutitas,Matricarias,Mayweeds, Crown,Pineapple Weeds,Weed, Pineapple,Weeds, Pineapple,chamomilla, Matricaria,matricarioide, Matricaria,recutitas, Chamomilla,recutitas, Matricaria