Molecular cloning of a novel trypsin-like serine protease (neurosin) preferentially expressed in brain.
A cDNA encoding a precursor for a novel serine protease (neurosin) was cloned from a cDNA library prepared from a human colon adenocarcinoma cell line, COLO 201. The sequence consisted of 155 bp 5' non-coding region and a 732 bp open reading frame which was followed by a 551 bp 3' non-coding region. The predicted protein consists of 244 amino acids which is possibly processed to an active enzyme of 223 amino acids that shows some similarity (< 30%) to other members of serine protease family. As found in other trypsin-like proteases, the enzyme contains the catalytic triad which is characterized as the essential amino acid residues for the activity. Northern blot analyses of the mRNA showed the strongest expression in brain followed by a lower but significant one in spleen. A construct of cDNA encoding chimeric protein that carries pro-sequence of trypsin II and putative mature neurosin starting from Leu22 was transfected to COS-1 cells. Successful production of the active neurosin was shown after treating the supernatant of the culture of the transfectants with enterokinase.
Yamashiro
K
K
Suntory Institute for Biomedical Research, Osaka, Japan.
Tsuruoka
N
N
Kodama
S
S
Tsujimoto
M
M
Yamamura
Y
Y
Tanaka
T
T
Nakazato
H
H
Yamaguchi
N
N
D000230:Adenocarcinoma; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D019556:COS Cells; D002460:Cell Line; D003001:Cloning, Molecular; D003110:Colonic Neoplasms; D005260:Female; D015723:Gene Library; D006801:Humans; D007610:Kallikreins; D008297:Male; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009928:Organ Specificity; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000737:chemistry; D014162:Transfection; D014357:Trypsin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D014407:Tumor Cells, Cultured
D011994:Recombinant Proteins; C103551:KLK6 protein, human; D007610:Kallikreins; D012697:Serine Endopeptidases; D014357:Trypsin