PURPOSE: To provide the subject new factor consisting of a protein isolated from pig pancreas, having a specified molecular weight and a specified amino acid sequence, having a protease activity, receiving an inhibitory effect by chymostatin, etc., and used, e.g. for therapy and prevention or osteopathic diseases such as osteoporosis.
CONSTITUTION: An acetone powder of pig pancreas is prepared and then extracted by using 0.1 M tris-hydrogen chloride buffer solution containing 2 wt.% NaCl. Acetone is subsequently added thereto till 30-60 wt.% concentration to obtain an acetone precipitated fraction. The resultant precipitated fraction is dialyzed by using deionized water and then desalted by ammonium sulfate salting out. The desalted crude extract is purified by an ion-exchange chromatography, thus obtaining the objective serum calcium-reducing factor capable of lowering the serum calcium level of a mouse in proportion to dosage, having 26500-28000 D (SDS-PAGE method) molecular weight and about 4.5 isoelectric point, having an N-terminal amino acid sequence of the formula and showing a protease activity receiving an inhibitory effect due to chymostatin and trypsin inhibitor and free from an inhibitory effect due to (4-amidinophenyl) methanesulfonyl fluoride, etc.
SAEKI TAKEYORI
TAKAOKA YOSHITO
TAKAOKA YOSHITO
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