Determination of phytochelatins and glutathione in phytoplankton from natural waters using HPLC with fluorescence detection

Kawakami, Silvia Keiko; Gledhill, Martha; Achterberg, Eric P.

doi:10.1016/j.trac.2005.06.005

Cited by 49 publications

(48 citation statements)

References 36 publications

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“…This peak was also observed in other MBrB-based derivatization studies (e.g., Thangavel et al, 2007;Minocha et al, 2008). Kawakami et al (2006) identified this peak as tetramethylbimane (Me 4 B) and reported that this compound was used during the synthesis of MBrB.…”

Section: Hplc Profile Of Thiol-containing Compoundssupporting

confidence: 66%

Reduced translocation of cadmium from roots is associated with increased production of phytochelatins and their precursors

Akhter

McGarvey

Macfie

2012

Journal of Plant Physiology

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SummaryCadmium (Cd) is a non-essential trace element and its environmental concentrations are approaching toxic levels, especially in some agricultural soils. Understanding how and where Cd is stored in plants is important for ensuring food safety. In this study, we examined two plant species that differ in the distribution of Cd among roots and leaves. Lettuce and barley were grown in nutrient solution under two conditions: chronic (4 week) exposure to a low, environmentally relevant concentration (1.0 µM) of Cd and acute (1 h) exposure to a high concentration (5.0 mM) of Cd. Seedlings grown in solution containing 1.0 µM CdCl 2 did not show symptoms of toxicity and, at this concentration, 77% of the total Cd was translocated to leaves of lettuce, whereas only 24% of the total Cd was translocated to barley leaves. We tested the hypothesis that differential accumulation of Cd in roots and leaves is related to differential concentrations of phytochelatins (PCs), and its precursor peptides. The amounts of PCs and their precursor peptides in the roots and shoots were measured using HPLC. Each of PC 2-4 was synthesized in the barley root upon chronic exposure to Cd and did not increase further upon acute exposure. In the case of lettuce, no PCs were detected in the root given either Cd treatment.The high amounts of PCs produced in barley root could have contributed to preferential retention of Cd in barley roots.

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Section: Hplc Profile Of Thiol-containing Compoundssupporting

confidence: 66%

Reduced translocation of cadmium from roots is associated with increased production of phytochelatins and their precursors

Akhter

McGarvey

Macfie

2012

Journal of Plant Physiology

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“…Because of their ubiquity but lower abundance, OSHs provide a greater opportunity for saturating the reactive sites, resulting in more reproducible quantitative labeling and less impact on the proteins. Numerous spectrometric methods for determining the OSHs derivatized by alkylation agents have been reported [4,5]. Although there are increasing numbers of commercially available thiol-reactive dyes, such as iodoacetamide or maleimide, the application of these dyes to proteomic studies still has some limitations.…”

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confidence: 99%

Counting sulfhydryls and disulfide bonds in peptides and proteins using mercurial ions as an MS-tag

Guo

Chen

Yang

et al. 2008

J. Am. Soc. Mass Spectrom.

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Organic mercurial compounds are the most specific and sensitive reagents for reaction with the sulfhydryl groups (OSHs) in peptides and proteins because of the strong mercury-sulfur affinity. Using the monofunctional organic mercury ion RHg ϩ as a mass spectrometry (MS)-tag has the advantages of reacting with one sulfhydryl group, offering definite mass shift, and especially stable and characteristic nonradioactive isotopic distribution. Mass spectrometric analysis of derivatized sulfhydryls in peptides/proteins is thus an alternative for precisely counting the number of sulfhydryl groups and disulfide bonds (OSOSO). Here the tags used include monomethylmercury chloride, monoethylmercury chloride, and 4-(hydroxymercuri) benzoic acid. The feasibility of this strategy is demonstrated using HPLC/ESI-MS to count OSHs and OSOSO in model peptides/proteins, i.e., glutathione, phytochelatins, lysozyme and ␤-lactoglobulin, which contain increasing OSHs and various OSOSO linkages. (J Am

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“…The powdered samples were extracted in 1.8 ml 5 mmol L −1 diethylene triamine pentaacetic acid (DTPA) containing 0.12 mmol L −1 HCl according to the method of Kawakami et al (2006), and then centrifuged at 18,000 × g at 4 • C for 10 min. The 250 L supernatant (or approximately diluted with 5 mmol L −1 DTPA containing 0.12 mmol L −1 HCl) or standards solutions were mixed with 200 mmol L −1 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer containing 5 mmol L −1 DTPA (pH 9.0, 650 L) and 20 mmol L −1 tris (2-carboxyethyl) phosphine (TCEP) (25 L).…”

Section: Internal Cadmium Quantificationmentioning

confidence: 99%

Phytochelatin synthesis in response to elevated CO2 under cadmium stress in Lolium perenne L.

Jia

Liao

et al. 2011

Journal of Plant Physiology

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Determination of phytochelatins and glutathione in phytoplankton from natural waters using HPLC with fluorescence detection

Cited by 49 publications

References 36 publications

Reduced translocation of cadmium from roots is associated with increased production of phytochelatins and their precursors

Reduced translocation of cadmium from roots is associated with increased production of phytochelatins and their precursors

Counting sulfhydryls and disulfide bonds in peptides and proteins using mercurial ions as an MS-tag

Phytochelatin synthesis in response to elevated CO2 under cadmium stress in Lolium perenne L.

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