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PMID: 3010323 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Isolation of a cDNA clone for the human lysosomal proteinase cathepsin B.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 9 ·1986-05-00 ·Pages 2909-13

Fong D, Calhoun DH, Hsieh WT, Lee B, Wells RD

Abstract

The cysteine proteinase cathepsin B is one member of the lysosomal acid hydrolases. Based on the peptide sequence of rat liver cathepsin B, an oligonucleotide mixture containing 128 different 17-mers was synthesized and used as a probe to screen adult and fetal human liver cDNA libraries. A recombinant clone with a 1540-nucleotide insert was identified from the fetal library, and DNA sequence analysis confirmed that this clone encodes human cathepsin B. The clone, designated pCB-1, has sequences for 81% of the coding region (for amino acid residues 50-252) together with approximately equal to 880 nucleotides of the 3' untranslated region of the mRNA. The DNA sequence also shows that the predicted carboxyl terminus of the coding sequence is longer than the mature protein by 6 amino acid residues. Southern blot analysis of restriction enzyme digests of human placental DNA revealed a simple pattern of hybridizing fragments using the cathepsin B coding sequence as probe. The result suggests that there is a single copy of cathepsin B gene per haploid genome.

MeSH Terms
Amino Acid Sequence Base Sequence Calpain/genetics Cathepsin B Cathepsins/analysis,genetics Cysteine Endopeptidases DNA/analysis,isolation & purification Endopeptidases/genetics Humans Isoenzymes/analysis Liver/analysis Lysosomes/enzymology Nucleic Acid Hybridization
Chemicals
Isoenzymes DNA Cathepsins Endopeptidases Calpain Cysteine Endopeptidases Cathepsin B
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fong D
Calhoun D H
Hsieh W T
Lee B
Wells R D
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44 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.9.2909
Published
1986-05-00
Pages
2909-13
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC323416
Subset
IM
Grants
NIGMS NIH HHS · GM-30822 · United States
Databases
GENBANK
M13230
External Links
PubMed source DOI: 10.1073/pnas.83.9.2909 PMC Full Text View journal details
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