Phylogenetic conservation of cysteine proteinases. Cloning and expression of a cDNA coding for human cathepsin S.
A 1.8-kilobase full-length cDNA of human cathepsin S, a lysosomal cysteine proteinase, has been isolated. The single long open reading frame encodes a polypeptide of 331 amino acids consisting of a 15-amino acid NH2-terminal signal peptide, a propeptide of 99 amino acids, and a mature polypeptide of 217 amino acids. The deduced amino acid sequence contains only one potential N-glycosylation site located in the propeptide. The NH2-terminal amino acid sequence of the mature polypeptide was confirmed by sequencing cathepsin S purified from human spleen. The cDNA detects a 1.9-kilobase transcript in poly(A)+ RNA from human fibroblasts. Expression of human cathepsin S in transfected baby hamster kidney cells resulted in up to more than 300-fold cathepsin S activity as compared to untransfected controls. In the expressing baby hamster kidney cells, human cathepsin S is transported to the lysosomes via the mannose 6-phosphate receptor pathway as shown by density gradient centrifugation, immunofluorescence, and detection of the 37-kDa cathepsin S precursor in the medium in the presence of NH4Cl. The deduced amino acid sequence of human cathepsin S exhibits a substantial degree of similarity with other human cysteine proteinases and papain indicating that they have a common ancestral gene and are members of a gene family.
Wiederanders
B
B
Institute of Physiological Chemistry, Martin Luther University, Halle, Federal Republic of Germany.
Brömme
D
D
Kirschke
H
H
von Figura
K
K
Schmidt
B
B
Peters
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002403:Cathepsins / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008969:Molecular Sequence Data; D010802:Phylogeny; D012313:RNA / Q000235:genetics; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D012313:RNA; D004247:DNA; D002403:Cathepsins; D003546:Cysteine Endopeptidases; C028217:cathepsin S