Glutathione S-Transferase Activity in Nontreated and CGA-154281- Treated Maize Shoots

Abstract Fast protein liquid chromatography (anion exchange) was used to separate glutathione S-transferase isozymes in nontreated etiolated maize shoots and those treated with the herbicide safener CGA -1542814-(dichloroacetyl)-3,4-dihydro-3-methyl-2 H-1 ,4-benzoxazine. Nontreated shoots contained isozymes active with the following substrates: trans-cinnamic acid (1 isozyme), atrazine (3 isozymes), 1-chloro-2,4-dinitrobenzene (1 isozyme), metolachlor (2 isozymes) and the sulfoxide derivative of S-ethyl dipropylcarbamothioate (2 isozymes). Pretreatment of shoots with the safener CGA -154281 (1 μM) had no effect on the activity of the isozymes selective for trans-cinnamic acid and atrazine but increased the activity of the constitutively-expressed isozymes that exhibit activity with 1-chloro-2,4-dinitrobenzene, metolachlor and the sulfoxide derivative of S-ethyl dipropylcarbamothioate. The safener pretreatment also caused the appearance of one new isozyme active with 1-chloro-2,4-dinitrobenzene and one new isozyme active with metolachlor. The results illustrate the complexity of glutathione S-transferase activity in etiolated maize shoots, and the selective enhancement of glutathione S-transferase isozymes by the safener CGA -154281.

Download Full-text

Tryptic phosphopeptides from whole casein. I. Preparation and analysis by fast protein liquid chromatography

Journal of Dairy Research ◽

10.1017/s0022029900029125 ◽

1989 ◽

Vol 56 (4) ◽

pp. 603-611 ◽

Cited By ~ 38

Author(s):

Marcel A. Juillerat ◽

Robert Baechler ◽

Raphael Berrocal ◽

Serge Chanton ◽

Jean-Claude Scherz ◽

...

Keyword(s):

Liquid Chromatography ◽

Anion Exchange ◽

Exchange Resin ◽

Anion Exchange Resin ◽

Gradient Elution ◽

Fast Protein Liquid Chromatography ◽

Parent Species ◽

Net Charge ◽

Phosphorylated Peptides ◽

Salt Gradient

SummaryTryptic phosphopeptides were obtained from whole bovine casein by chromatography on the anion exchange resin QAE-Sephadex A 25. Salt gradient elution of the column allowed separation of non-phosphorylated peptides from phosphorylated species. The preparations obtained contained at least seven distinct phosphopeptides of which the following casein fragments were identified: αs1(43–58):2P, αs1(59–79): 5P, αs2(46–70): 4P, β(1–28): 4P, β(2–28): 4P, and β(33–48): 1P. Fast protein liquid chromatography (FPLC) on Mono Q HR 5/5 resin showed that the phosphopeptides were eluted in the same order as from the QAE-Sephadex resin. However, on the analytical column HR 5/5 the fragments αs1(59–79): 5P and β(2–28): 4P, having the same net charge under the conditions of chromatography, co-eluted, whereas they were at least partly separated on the preparative column HR 16/10. Following enzymic dephosphorylation, the peptides eluted at lower salt strength in the gradient. FPLC on Mono Q resin thus permitted dephosphorylation to be monitored and intermediates between the parent species and the fully dephosphorylated peptide to be identified.

Download Full-text

Applications of fast protein liquid chromatography TM in the separation of plasma proteins in urine and cerebrospinal fluid.

Clinical Chemistry ◽

10.1093/clinchem/29.9.1635 ◽

1983 ◽

Vol 29 (9) ◽

pp. 1635-1640 ◽

Cited By ~ 9

Author(s):

E H Cooper ◽

R Turner ◽

E A Johns ◽

H Lindblom ◽

V J Britton

Keyword(s):

Cerebrospinal Fluid ◽

Liquid Chromatography ◽

Anion Exchange ◽

Plasma Proteins ◽

Fast Protein Liquid Chromatography ◽

Rapid Separation ◽

Anion Exchange Column ◽

Exchange Column ◽

Urine Specimens ◽

Bence Jones Proteins

Abstract Fast Protein Liquid Chromatography TM (FPLC), in which an anion-exchange column is used, provides rapid separation and reproducible profiling of the plasma proteins in urine and cerebrospinal fluid (CSF). Chromatographic separation of the proteins takes 1 h for urine specimens and 45 min for CSF. The elution sequence from the anion-exchange column is similar to the electrophoretic mobility. Individual proteins have the same retention times independently of which type of specimen is used. The elution characteristics of 21 plasma proteins have been identified. We illustrated some applications of this system, including the profiling of tubular protein-uria, the isolation of Bence Jones proteins from urine, and the investigation of hemoglobin-derived products in the CSF.

Download Full-text