Molecular cloning of complementary DNA encoding one of the human pancreatic protease E isozymes.
We have cloned a DNA from a human pancreatic cDNA library using a cloned rat pancreatic elastase 1 cDNA as a probe, and determined its nucleotide sequence. This cDNA contains a coding region of 810 nucleotides which encodes a 270-amino-acid protein. The deduced amino acid sequence shows less than 60% homologies with rat and porcine pancreatic elastase 1, although its substrate binding region is homologous with those of the above elastases 1. When this deduced amino acid sequence was compared with known amino acid sequences of pancreatic proteases other than elastases, it was found to contain an amino acid sequence which was highly homologous with the N-terminal amino acid sequence of porcine pancreatic protease E. We also purified human pancreatic protease E isozymes from human pancreatic juice, and determined their N-terminal amino acid sequences. One of the isozymes does not hydrolyze elastin but does hydrolyze a synthetic substrate. Endoglycosidase F digests glycoside bonds of the isozyme. These results suggest that the cDNA cloned by us corresponded to one of the human protease E isozymes.
Shirasu
Y
Y
Pharmaceutical Laboratory, Kirin Brewery Co., Ltd., Gunma.
Takemura
K
K
Yoshida
H
H
Sato
Y
Y
Iijima
H
H
Shimada
Y
Y
Mikayama
T
T
Ozawa
T
T
Ikeda
N
N
Ishida
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D012313:RNA / Q000302:isolation & purification; D012697:Serine Endopeptidases / Q000235:genetics*
D007527:Isoenzymes; D012313:RNA; D004247:DNA; D012697:Serine Endopeptidases; C028734:protease E