Purification of two equine pepsinogens by use of high-performance liquid chromatography.

The research article discusses the purification and characterization of two types of pepsinogens located in the gastric mucosa of horses, with the hope of using this information as a basis for diagnosing equine gastric ulcers in the future.

Methodology

• The research team collected stomachs from two healthy horses post-mortem. The collected stomachs were immediately chilled and then stored at -48 degrees Celsius.
• After a slow thawing process, the gastric mucosa was scrapped off while it was immersed in a 0.1M potassium phosphate solution at a pH of 7.4 and at a temperature of 4 degrees Celsius. This mixture was then homogenized.
• The filtered extract was then subjected to anion-exchange chromatography. The fractions that were found to contain pepsin or pepsinogen were then further subjected to chromatography.
• Each individual fraction was tested for its content of pepsinogen or pepsin by observing their proteolytic activities at pH 2 and 3, respectively.
• These fractions were then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis to validate pepsinogens and pepsin’s molecular weight.

Results

• The researchers were able to purify two types of pepsinogens and at least one pepsin from the equine gastric mucosa.

Conclusions

• The gastric mucosa of horses was established to contain two types of pepsinogens based on molecular mass.
• The results from this study provide valuable insights which can aid in the development of testing procedures such as ELISA or radioimmunoassay for diagnosing horse gastric ulcerations in the future.