Immobilized-metal-ion affinity chromatography as a tool for the qualitative study of pepsinogen phosphorylation.
Affinity chromatography on immobilized Fe(3+) ions--immobilized-metal-ion affinity chromatography (IMAC) method--was used for the determination of pepsin and pepsinogen phosphorylation. IMAC is a very powerful method for detailed studies of proteins. Dephosphorylation of the pepsinogens and pepsins has no effect on their proteolytic ability. For this reason, the determination of proteolytic activity was used for the detection of pepsinogen (pepsin) presence in the collected fractions as a very suitable and specific method. Pepsins and their zymogens probably have the same amounts of phosphate ions in their molecule. The exact definition of conditions is very important for the prepurification of the proteinases and for their analysis.
Kucerová
Z
Z
1st Faculty of Medicine, Institute of Pathophysiology, Charles University, U nemocnice 5, 12853 Prague 2, Czech Republic. zkuce@lf1.cuni.cz
Majercaková
P
P
D000595:Amino Acid Sequence; D002021:Buffers; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D005290:Ferric Compounds; D005753:Gastric Mucosa / Q000201:enzymology*; D006801:Humans; D007202:Indicators and Reagents; D008024:Ligands; D008670:Metals; D009842:Oligopeptides; D010434:Pepsin A / Q000378:metabolism; D020116:Pepsinogen A / Q000378:metabolism*; D010766:Phosphorylation
D002021:Buffers; D005290:Ferric Compounds; D007202:Indicators and Reagents; D008024:Ligands; D008670:Metals; D009842:Oligopeptides; D020116:Pepsinogen A; D010434:Pepsin A