Functional expression of human leukocyte elastase (HLE)/medullasin in eukaryotic cells.
We have cloned a full length cDNA for human leukocyte elastase (HLE, EC 3.4.21.37)/medullasin from the cDNA library of human leukemic cell line, ML3. Recombinant plasmid for the expression of HLE cDNA in eukaryotic cells was constructed in which HLE cDNA was fused in a frame to a leader sequence of human interleukin-2 (IL-2). COS-1 cells, transfected with the plasmid, secreted fusion protein consists of N-terminal 8 amino acid (aa) residues of human IL-2 and 238 aa residues of HLE. As the fusion protein was designed to be connected through lysine residue, elastase activity was generated after digestion of the fusion protein with lysyl-endopeptidase.
Okano
K
K
Basic Research Laboratories, Toray Industries Inc., Kamakura, Japan.
Aoki
Y
Y
Shimizu
H
H
Naruto
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000235:genetics; D007962:Leukocytes / Q000201:enzymology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010196:Pancreatic Elastase / Q000235:genetics*; D010957:Plasmids; D015183:Restriction Mapping; D012697:Serine Endopeptidases / Q000235:genetics*; D014162:Transfection
D007376:Interleukin-2; D015345:Oligonucleotide Probes; D004247:DNA; D012697:Serine Endopeptidases; C034869:medullasin; D010196:Pancreatic Elastase