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Purification of two equine pepsinogens by use of high-performance liquid chromatography
A I Sayegh1, N V Anderson, J W Harding
1Department of Clinical Sciences, Kansas State University, Manhattan 66506, USA.
Objectives:
To purify and characterize pepsinogens in equine gastric mucosa.
Sample Population:
Stomachs collected from 2 healthy horses at necropsy.
Procedure:
After collection, stomachs were placed immediately in ice before storage at -48 C. After slow thawing, the mucosa was scraped off while the tissue was immersed in 0.1M potassium phosphate (pH 7.4) at 4 C, then was homogenized. The filtered extract was subjected to anion-exchange chromatography. Fractions that were found to contain pepsin or pepsinogen were further chromatographed. Individual fractions were tested for pepsinogen or pepsin content by monitoring proteolytic activity at pH 2 and 3, respectively. Fractions from all columns were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to confirm molecular weight of pepsinogens and pepsin.
Results:
Two pepsinogens and at least 1 pepsin were purified from equine gastric mucosa.
Conclusions:
On the basis of molecular mass, equine gastric mucosa contains 2 pepsinogens.
Clinical Relevance:
Results of this study will enable future development of an ELISA or radioimmunoassay for use in the diagnosis of equine gastric ulceration.
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