Rapid chromatographic purification of dipeptidyl peptidase IV in human submaxillary gland.

Pure dipeptidyl peptidase IV (X-prolyl dipeptidyl aminopeptidase), which did not contain aminopeptidase activity at all, was rapidly prepared from the human submaxillary gland by chromatography with concanavalin A-Sepharose and Gly-Pro-NH-(CH2)6-NH-Sepharose. The entire purification took only 3 days. Aminopeptidase, which was very difficult to separate from dipeptidyl peptidase IV by various chromatographic procedures, could be completely removed by chromatography with Gly-Pro-NH-(CH2)6-NH-Sepharose. On SDS gel electrophoresis the purified enzyme gave a single band with a molecular weight of 116,000. The apparent molecular weight of the enzyme was estimated to be 225,000 by gel filtration. Therefore, the enzyme consists of two identical subunits. It did not hydrolyze Ala p-nitroanilide at all, but the hydrolysis of the p-nitroanilides of Gyl-Pro, Lys-Pro and Arg-Pro at pH 8.0 was nearly specific.