Analysis of phytochelatin-cadmium complexes from plant tissue culture using nano-electrospray ionization tandem mass spectrometry and capillary liquid chromatography/electrospray ionization tandem mass spectrometry.
Phytochelatins (PCs, also known as class III metallothioneins), a family of sulfhydryl-rich peptides with the formula (gamma-GluCys)(n)Gly(Pc(n), n = 2-11), are induced in plants, yeast and fungi exposed to heavy metals, and are thought to detoxify metals by forming PC- metal complexes. Although PCs have been detected, PC- metal complexes have not been well characterized. In this work, nano-electrospray ionization tandem mass spectrometry (nano-ESI-MS/MS) and capillary liquid chromatography/electrospray ionization tandem mass spectrometry (capillary LC/ESI-MS/MS) methods were used to analyze PC - Cd complexes isolated from Datura innoxia, also known as Jimsonweed, cell culture exposed to Cd. With nano-ESI-MS/MS and capillary LC/ESI-MS/MS we could simultaneously detect the presence of PCs and PC - Cd complexes from plant cell extracts, unambiguously identify these species and elucidate the nature of individual PC - Cd complexes. Phytochelatins with n = 3-6 were detected, as were PC - Cd complexes with PC(3), PC(4) and PC(5). This is the first study to report the size and nature of native PC - Cd complexes from plant tissue samples. These results demonstrate that the direct analysis of plant extracts using nano-ESI-MS/MS and capillary LC/ESI-MS/MS methods is simple and sensitive to the range of PCs and PC - Cd complexes in plants. Hence these methods open up new opportunities for further quantitative analysis of PCs and PC - metal complexes in cell culture and plant systems to understand the relationship between the biosynthesis of these compounds and metal tolerance.
Yen
T Y
TY
Biomedical Research Mass Spectrometry Facility, Department of Chemistry and Biochemistry, San Francisco State University, San Francisco, California 94132-4163, USA.
Villa
J A
JA
DeWitt
J G
JG
D000595:Amino Acid Sequence; D002104:Cadmium / Q000378:metabolism*; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods; D005978:Glutathione; D013058:Mass Spectrometry / Q000379:methods; D008667:Metalloproteins / Q000737:chemistry* / Q000378:metabolism*; D054811:Phytochelatins; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism*; D010944:Plants / Q000378:metabolism; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods
D008667:Metalloproteins; D010940:Plant Proteins; D002104:Cadmium; D054811:Phytochelatins; D005978:Glutathione