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On the Activation of Bovine Chymotrypsinogen A. Preparation of Alanine-Neochymotrypsinogen and Its Activation to Alpha-Chymotrypsin

On the Activation of Bovine Chymotrypsinogen A. Preparation of Alanine-Neochymotrypsinogen and Its Activation to Alpha-Chymotrypsin

S K Sharma, T R Hopkins

Biochim Biophys Acta. 1978 Dec 8;527(2):465-71.

PMID: 569499

Abstract:

Alanine-neochymotrypsinogen was prepared by incubating 20 parts bovine pancreas chymotrypsinogen A with one part alpha-chymotrypsin in a solution containing 1 M (NH4)2SO4, 0.1 M sodium acetate, 0.05 M Tris buffer (pH 8.0) and 0.5 mg/ml soybean trypsin inhibitor. Optimal yields of NH2-terminal alanine were obtained after 60 h incubation at 4 degrees C. Ala-neochymotrypsinogen was isolated from the reaction mixture by affinity chromatography and ion-exchange chromatography on carboxymethyl-cellulose. As expected, the purified preparation was enzymatically inactive and, compared to chymotrypsinogen, had one additional NH2-terminal group identified as alanine. Ala-neochymotrypsinogen was activated by incubating with trypsin at a zymogen : trypsin ratio of 30 : 1 in 0.1 M phosphate buffer, pH 7.6 at 4 degrees C for 1 h. The fully active, stable species was identified as alpha-chymotrypsin.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP9035750	α-Chymotrypsinogen A from bovine pancreas		9035-75-0	Price

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