Journal article

PURIFICATION AND CLONING OF A NOVEL SERINE PROTEASE, RNK-MET-1, FROM THE GRANULES OF A RAT NATURAL-KILLER-CELL LEUKEMIA

MJ SMYTH, T WILTROUT, JA TRAPANI, KS OTTAWAY, R SOWDER, LE HENDERSON, CM KAM, JC POWERS, HA YOUNG, TJ SAYERS

Journal of Biological Chemistry | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC | Published : 1992

Abstract

We have purified a 30-kDa serine protease (designated RNK-Met-1) from the granules of the rat large granular lymphocyte leukemia cell line (RNK-16) that hydrolytically cleaves model peptide substrates after methionine, leucine, and norleucine (Met-ase activity). Utilizing molecular sieve chromatography, heparin-agarose, chromatography, and reverse-phase high pressure liquid chromatography, RNK-Met-1 was purified to homogeneity and 25 NH2-terminal amino acids were sequenced. By using the polymerase chain reaction, oligonucleotide primers derived from amino acids at position 14-25 and from a downstream active site conserved in other serine protease genes were used to generate a 534-base pair c..

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