Abstract
Abstract Pepsinogen has been purified from the fundic mucosa of the dog. The preparation is homogeneous by chromatography, amino terminal analysis, ultracentrifugation, electrophoresis on cellulose acetate strips, and disc electrophoresis. Analysis indicates the presence of 352 amino acid residues, at least 12 moles of carbohydrate including some amino sugars, and 1 mole of organic phosphate per mole of protein. The amino terminus is Ala-Ile-. Canine, porcine, and bovine pepsinogens are similar in size, in the large number of acidic and small number of basic residues, and in the number of aromatic residues and cystine. The main differences are in the number of hydroxy amino acids and methionines; canine pepsinogen with 8 methionine residues is unlike the other mammalian zymogens, but resembles dogfish and chicken pepsinogens. The molecular weight of canine pepsinogen is 39,900 by amino acid and carbohydrate content and 41,667 by ultracentrifugation; the sedimentation coefficient, s20,ω, is 3.38 S and the diffusion coefficient, D20,ω, is 7.37 x 10-7 cm2 per sec
Highlights
The amino terminal group was identified by thin layer chromatography in Systems I and II of Gros and Labouesse [19]
The presence of amino sugars was established by amino acid analysis [22]
The total number of amino acids is quite similar for the three pepsinogens
Summary
Amino acid analysis was carried out on a Beckman-Spinco model 120B analyzer, essentially as described byMoore and Stein [12]; the details were presented in a previous paper [5]. Amino acid analysis was carried out on a Beckman-Spinco model 120B analyzer, essentially as described by. The amino terminal group at each step was measured as the PTH3-derivative [15] and identified by thin layer chromatography on silica gel plates [16, 17]. Was used for amino terminal analysis of the zymogen. The amino terminal group was identified by thin layer chromatography in Systems I and II of Gros and Labouesse [19]. Total carbohydrate was determined by the phenol-sulfuric acid method [20] after deamination as described by Lee and Montgomery [21] ; the standard was an equimolar mixture of glucose and galactosamine. Carbohydrate content, exclusive of amino sugars, \yas determined directly by the phenol-sulfuric acid method [20] with a glucose standard.
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