Prefractionation of Intact Proteins by Reversed-Phase and Anion-Exchange Chromatography for the Differential Proteomic Analysis ofSaccharomyces cerevisiae

2012 ◽  
Vol 12 (2) ◽  
pp. 626-636 ◽  
Author(s):  
Jordan T. Stobaugh ◽  
Kaitlin M. Fague ◽  
James W. Jorgenson
Keyword(s):  
Anion Exchange ◽  
Proteomic Analysis ◽  
Anion Exchange Chromatography ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
Intact Proteins

Rapid analysis for iodotyrosines and iodothyronines in thyroglobulin by reversed-phase liquid chromatography.

1979 ◽  
Vol 25 (10) ◽  
pp. 1757-1760 ◽  
Author(s):  
N M Alexander ◽  
M Nishimoto
Keyword(s):  
Anion Exchange ◽  
Anion Exchange Chromatography ◽  
Reversed Phase ◽  
Exchange Chromatography ◽  
Chart Recorder ◽  
Chromatographic Procedure ◽  
Phase Liquid ◽  
Flow Through ◽  
Sequential Digestion ◽  
Liquid Chromatographic

Abstract We describe a 10-min reversed-phase "high-pressure' liquid-chromatographic procedure for measuring tyrosine, monoiodotyrosine, diiodotyrosine, 3,5-diiodothyronine, 3,5,3'-triiodothyronine, 3,3',5'-triiodothyronine, and thyroxine. Resolution and quantitation of a mixture of these amino acids were excellent on LiChrosorb (Altex) RP-8 with isocratic elution (1.5 mL/min) with acetonitrile/water/glacial acetic acid (50/49/1 by vol). As little as 100 ng of each iodoamino acid could be detected and quantitated with a conventional 1-cm, flow-through spectrophotometric (254-nm) detector coupled to a 10-mV strip-chart recorder. Analyses for monoiodotyrosine, diiodotyrosine, 3,5,3'-triiodothyronine, and thyroxine in hog and beef thyroglobulin hydrolysates (sequential digestion with pronase and aminopeptidase) agreed well with results by anion-exchange chromatography and by competitive radioassays. To prevent interference by tryptophan in the analysis for diiodotyrosine, we batch-separated the iodoamino acids by anion-exchange chromatography before the procedure. The procedure we describe seems generally useful for detection and quantitation of thyroid hormones, thyroid hormone metabolites, and iodotyrosines.

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