Biol. Chem. Hoppe-Seyler Vol. 373, pp. 367-373, July 1992
EIKI KOMINAMI, KAZUMI ISHIDO, DAISAKU MUNO AND NOBOURU SATO
Department of Biochemistry, Juntendo University School of Medicine, 2-1-1, Kongo, Bunkyo-ku, Tokyo 113, Japan Summary
A cDNA for rat cathepsin C (dipeptidylaminopeptidase I) was isolated. The encoded protein is composed of the signal peptide of 28 residues, the propeptide of 201 residues and the mature enzyme region of 233 residues. The amino acid sequence of the mature enzyme region has 39.5 to 30.5 % identity to other papain family proteinases. Cathepsin C is, therefore, belongs to papain family, although its propeptide region is much longer than those of other cysteine proteinases and show no significant sequence similarity to any other cysteine proteinase. The mRNA and protein for cathepsin C are broadly distributed in rat tissues, but the relative proportions of cathepsin C and other cysteine proteinases are found to vary from tissue to tissue. Introduction The primary structures of lysosomal endopeptidases such as cathepsins B,H,L,S and D have been determined from varous mammals by both protein sequencing [1-3] and cDNA cloning experiments [4-7]. However, no exopeptidases have been sequenced except cathepsin A (lysosomal carboxypeptidase A), which was identified as lysosomal " protective protein " [8]. Among the lysosomal exopeptidases, dipeptidyl peptidase I is the one that has been most extremely purified and characterized. This enzyme, originally called cathepsin C, was discovered by Gutman and Fruton [9] when extracts of porcine kidney were found to catalyze the hydrolysis of Gly-Phe-NH2. Later, this enzyme was characterized as a cysteine-dependent Ser-Tyr-j8-naphtylamidase with an absolute or near absolute chloride requirement [10]. Its action was found to be so non-specfic as to catalize the removal of several dipeptides in succession from N-terminus of large hormonal polypeptides [11]. But further characterization including primary structure and cellular function has not been studied so far. In this report we present the amino acid sequence and tissue distribution of rat cathepesin C.
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368
E. Kominami, K. Ishido, D. Muno and N. Sato
Vol. 373 (1992)
Cathepsin C is a member of papain family cDNA clones for cathepsn C were isolated from a rat kidney cDNA library [12]. The nucleotide sequence of the cDNA for rat cathepsin C comprises 1842 bp containing 49 bp in the 5'-noncoding region, 1389 bp in the coding region, and 404 bp in the 3'-non coding region. Preprocathepsin C comprises 462 amino acid residues containing three functional domains. The 28 aminoterminal residues corespond to the signal peptide, the next 201 amino acid residues correspond to the proregion (Fig.l). The 233 carboxyterminal residues correspond to the 1
τ
50 100 150
* 300 QESCGSCYSFAS I GMLEAR I R I LTNNSQTP I LSPQEWSCSP YAQGCDGG 350 FP YL I AGKY AQDFGVVEENCFP YTATDAPCKPKENCLRY YSSE Υ Υ YVGGF 400 T YGGCNEALMKLELVKHGPMAVAFEVHDDFLHYHSGIYHHTGLSDPFNPFE * * 450 LTNHAVLLVGYGKDPVTGLDYWIVKNSWGSQWGESGYFRIRRGTDECAIE SIAMAAIPIPKL
Fig.l. Amino acid sequence of rat preprocathepsin C deduced from nucleotide sequence. Arrows indicate possible NH2 termini of the propeptide, the heavy chain and the light chains of the mature emzyme, respectively. The underlines indicate possible glycosylation sites. Asterics indicate the active cysteine, histidine and aspargine residues. mature region. In the mature enzyme region, the amino acid sequence of cathepsin C shares 39.5, 35.1, 30.1 and 33.3 % identity with those of cathepsins H, L, B and papain, respectively. The amino acid residues that form part of the active site of the enzyme are highly conserved. Especially, the amino acids involved in catalysis, e.g. Cys25 and His159, as well as Gin19, Asn175, Trp177 (papain numbering) are all retained in cathepsin C (Fig.2). N-terminal amino acids of the most majority of mature cysteine proteinases can be aligned exactly with one another, the second amino acid being proline. This rule is retained in cathepsin C. Thus, cathepsin C belongs to the papain family. The tyrosine residue next to the active site cysteine in cathepsin C is a substitution for the tryptophan residue that is well conserved among
Brought to you by | Purdue University Libraries Authenticated Download Date | 5/22/15 8:36 PM
Vol. 373 (1992)
Primary Structure andTissue Distribution of Cathepsin C
369
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EIKI KOMINAMI, KAZUMI ISHIDO, DAISAKU MUNO AND NOBOURU SATO
Department of Biochemistry, Juntendo University School of Medicine, 2-1-1, Kongo, Bunkyo-ku, Tokyo 113, Japan Summary
A cDNA for rat cathepsin C (dipeptidylaminopeptidase I) was isolated. The encoded protein is composed of the signal peptide of 28 residues, the propeptide of 201 residues and the mature enzyme region of 233 residues. The amino acid sequence of the mature enzyme region has 39.5 to 30.5 % identity to other papain family proteinases. Cathepsin C is, therefore, belongs to papain family, although its propeptide region is much longer than those of other cysteine proteinases and show no significant sequence similarity to any other cysteine proteinase. The mRNA and protein for cathepsin C are broadly distributed in rat tissues, but the relative proportions of cathepsin C and other cysteine proteinases are found to vary from tissue to tissue. Introduction The primary structures of lysosomal endopeptidases such as cathepsins B,H,L,S and D have been determined from varous mammals by both protein sequencing [1-3] and cDNA cloning experiments [4-7]. However, no exopeptidases have been sequenced except cathepsin A (lysosomal carboxypeptidase A), which was identified as lysosomal " protective protein " [8]. Among the lysosomal exopeptidases, dipeptidyl peptidase I is the one that has been most extremely purified and characterized. This enzyme, originally called cathepsin C, was discovered by Gutman and Fruton [9] when extracts of porcine kidney were found to catalyze the hydrolysis of Gly-Phe-NH2. Later, this enzyme was characterized as a cysteine-dependent Ser-Tyr-j8-naphtylamidase with an absolute or near absolute chloride requirement [10]. Its action was found to be so non-specfic as to catalize the removal of several dipeptides in succession from N-terminus of large hormonal polypeptides [11]. But further characterization including primary structure and cellular function has not been studied so far. In this report we present the amino acid sequence and tissue distribution of rat cathepesin C.
Brought to you by | Purdue University Libraries Authenticated Copyright © by Walter de Gruyter & Co · Berlin · New York Download Date | 5/22/15 8:36 PM
368
E. Kominami, K. Ishido, D. Muno and N. Sato
Vol. 373 (1992)
Cathepsin C is a member of papain family cDNA clones for cathepsn C were isolated from a rat kidney cDNA library [12]. The nucleotide sequence of the cDNA for rat cathepsin C comprises 1842 bp containing 49 bp in the 5'-noncoding region, 1389 bp in the coding region, and 404 bp in the 3'-non coding region. Preprocathepsin C comprises 462 amino acid residues containing three functional domains. The 28 aminoterminal residues corespond to the signal peptide, the next 201 amino acid residues correspond to the proregion (Fig.l). The 233 carboxyterminal residues correspond to the 1
τ
50 100 150
* 300 QESCGSCYSFAS I GMLEAR I R I LTNNSQTP I LSPQEWSCSP YAQGCDGG 350 FP YL I AGKY AQDFGVVEENCFP YTATDAPCKPKENCLRY YSSE Υ Υ YVGGF 400 T YGGCNEALMKLELVKHGPMAVAFEVHDDFLHYHSGIYHHTGLSDPFNPFE * * 450 LTNHAVLLVGYGKDPVTGLDYWIVKNSWGSQWGESGYFRIRRGTDECAIE SIAMAAIPIPKL
Fig.l. Amino acid sequence of rat preprocathepsin C deduced from nucleotide sequence. Arrows indicate possible NH2 termini of the propeptide, the heavy chain and the light chains of the mature emzyme, respectively. The underlines indicate possible glycosylation sites. Asterics indicate the active cysteine, histidine and aspargine residues. mature region. In the mature enzyme region, the amino acid sequence of cathepsin C shares 39.5, 35.1, 30.1 and 33.3 % identity with those of cathepsins H, L, B and papain, respectively. The amino acid residues that form part of the active site of the enzyme are highly conserved. Especially, the amino acids involved in catalysis, e.g. Cys25 and His159, as well as Gin19, Asn175, Trp177 (papain numbering) are all retained in cathepsin C (Fig.2). N-terminal amino acids of the most majority of mature cysteine proteinases can be aligned exactly with one another, the second amino acid being proline. This rule is retained in cathepsin C. Thus, cathepsin C belongs to the papain family. The tyrosine residue next to the active site cysteine in cathepsin C is a substitution for the tryptophan residue that is well conserved among
Brought to you by | Purdue University Libraries Authenticated Download Date | 5/22/15 8:36 PM
Vol. 373 (1992)
Primary Structure andTissue Distribution of Cathepsin C
369
50
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