Chromatographic separation of pepsins from human gastric juice

Abstract

Acid gastric juices have been fractionated on DEAE (diethylaminoethyl)-cellulose to yield three pepsin fractions, which have the same chromatographic mobility as the three pepsin fractions previously described in acidified whole gastric mucosal extracts. The acid gastric juices included specimens obtained from healthy individuals and from patients with peptic ulcer and gastritis. Gastric juice from one patient with pernicious anemia was shown to contain only pepsinogen I. A satisfactory chromatographic separation of pepsins is possible from as little as 4 ml of gastric juice.

Pepsin 5 in gastric juice: determination and relationship to the alkali-stable peptic activity.

“…Turner et al (1967) observed that the 4pepsin 1' of Seijffers et al (1964) was. stable at pH 70 and showed only a slow loss of activity between pH 7 0 and pH 7 5, whereas pepsins IIA, IIB, and III showed a rapid loss of proteolytic activity at pH 70.…”

Pepsin secretion in man after Histalog stimulation

“…0.1 M acetate buffer (pH 5 3) was made from 0-1 M sodium acetate and 0.1 M acetic acid. Appropriate amounts of sodium chloride, as described below, were added for gradientelution chromatography (Seijffers et al, 1964).…”

“…DiscussionThe chromatographic technique gave reasonably reproducible results(Seijffers et al, 1964) but temperature control was critical to obtain satisfactory separation of the three pepsin fractions. However, even with satisfactory chromatograms it was not possible to determine the proportions…”

“…The second specimen of basal secretion and all post-Histalog specimens of gastric juice were analysed by chromatography on a column of DEAE-cellulose 2 x 19.5 cm prepared as described by Seijffers et al (1964). An amount of each specimen containing 73.5 units of peptic activity was loaded on to each column.…”

“…It now appears that human gastric mucosa contains at least seven distinguishable proteinase proenzymes (Turner, Mangla, Samloff, Miller, and Segal, 1970) which are the precursors of a number of pepsins which are found in the gastric juice. Although, as yet, it is not clear how many individual enzymes result from the activation of these zymogens, and no method has yet been developed for the quantitative measurement of each individually, Seijffers, Segal, and Miller (1964) were able to separate human pepsin into three distinct fractions which could be measured quantitatively by chromatography on DEAE-cellulose. These fractions were termed 'pepsin I', 'pepsin II', and 'pepsin III'.…”