Quantification of phytochelatins in Chlamydomonas reinhardtii using ferrocene-based derivatization.
A method for the identification and quantification of canonic and isoforms of phytochelatins (PCs) from Chlamydomonas reinhardtii was developed. After disulfide reduction with tris(2-carboxyethyl)phosphine (TCEP) PCs were derivatized with ferrocenecarboxylic acid (2-maleimidoyl)ethylamide (FMEA) in order to avoid oxidation of the free thiol functions during analysis. Liquid chromatography (LC) coupled to electrospray mass spectrometry (ESI-MS) and inductively coupled plasma-mass spectrometry (ICP-MS) was used for rapid and quantitative analysis of the precolumn derivatized PCs. PC(2-4), CysGSH, CysPC(2-4), CysPC(2)desGly, CysPC(2)Glu and CysPC(2)Ala were determined in the algal samples depending on the exposure of the cells to cadmium ions.
Martin-Luther-Universität Halle-Wittenberg, Institut für Biochemie und Biotechnologie, Ökologische und Pflanzen-Biochemie, Kurt-Mothes-Straße 3, Halle (Saale), Germany.
Anja
A
Bräutigam
Susanne
S
Bomke
Thorben
T
Pfeifer
Uwe
U
Karst
Gerd-Joachim
GJ
Krauss
Dirk
D
Wesenberg
D016825:Chlamydomonas reinhardtii / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005296:Ferrous Compounds / Q000378:metabolism*; D000075163:Metallocenes; D010084:Oxidation-Reduction; D054811:Phytochelatins / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
D005296:Ferrous Compounds; D000075163:Metallocenes; D054811:Phytochelatins; C004998:ferrocene