Isolation of a cDNA clone for the human lysosomal proteinase cathepsin B.
Identifieur interne : 004E08 ( Main/Merge ); précédent : 004E07; suivant : 004E09Isolation of a cDNA clone for the human lysosomal proteinase cathepsin B.
Auteurs : D. Fong ; D H Calhoun ; W T Hsieh ; B. Lee ; R D WellsSource :
- Proceedings of the National Academy of Sciences of the United States of America [ 0027-8424 ] ; 1986.
Descripteurs français
- KwdFr :
- ADN (analyse), ADN (isolement et purification), Calpain (génétique), Cathepsine B, Cathepsines (analyse), Cathepsines (génétique), Cysteine endopeptidases, Endopeptidases (génétique), Foie (analyse), Humains, Hybridation d'acides nucléiques, Isoenzymes (analyse), Lysosomes (enzymologie), Séquence d'acides aminés, Séquence nucléotidique.
- MESH :
- analyse : ADN, Cathepsines, Foie, Isoenzymes.
- enzymologie : Lysosomes.
- génétique : Calpain, Cathepsines, Endopeptidases.
- isolement et purification : ADN.
- Cathepsine B, Cysteine endopeptidases, Humains, Hybridation d'acides nucléiques, Séquence d'acides aminés, Séquence nucléotidique.
English descriptors
- KwdEn :
- Amino Acid Sequence, Base Sequence, Calpain (genetics), Cathepsin B, Cathepsins (analysis), Cathepsins (genetics), Cysteine Endopeptidases, DNA (analysis), DNA (isolation & purification), Endopeptidases (genetics), Humans, Isoenzymes (analysis), Liver (analysis), Lysosomes (enzymology), Nucleic Acid Hybridization.
- MESH :
- chemical , analysis : Cathepsins, DNA, Isoenzymes.
- chemical , genetics : Calpain, Cathepsins, Endopeptidases.
- chemical , isolation & purification : DNA.
- analysis : Liver.
- enzymology : Lysosomes.
- Amino Acid Sequence, Base Sequence, Cathepsin B, Cysteine Endopeptidases, Humans, Nucleic Acid Hybridization.
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.
DOI: 10.1073/pnas.83.9.2909
PubMed: 3010323
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pubmed:3010323Le document en format XML
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<author><name sortKey="Hsieh, W T" sort="Hsieh, W T" uniqKey="Hsieh W" first="W T" last="Hsieh">W T Hsieh</name>
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<author><name sortKey="Lee, B" sort="Lee, B" uniqKey="Lee B" first="B" last="Lee">B. Lee</name>
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<term>Base Sequence</term>
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<term>Cathepsins (analysis)</term>
<term>Cathepsins (genetics)</term>
<term>Cysteine Endopeptidases</term>
<term>DNA (analysis)</term>
<term>DNA (isolation & purification)</term>
<term>Endopeptidases (genetics)</term>
<term>Humans</term>
<term>Isoenzymes (analysis)</term>
<term>Liver (analysis)</term>
<term>Lysosomes (enzymology)</term>
<term>Nucleic Acid Hybridization</term>
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<term>ADN (isolement et purification)</term>
<term>Calpain (génétique)</term>
<term>Cathepsine B</term>
<term>Cathepsines (analyse)</term>
<term>Cathepsines (génétique)</term>
<term>Cysteine endopeptidases</term>
<term>Endopeptidases (génétique)</term>
<term>Foie (analyse)</term>
<term>Humains</term>
<term>Hybridation d'acides nucléiques</term>
<term>Isoenzymes (analyse)</term>
<term>Lysosomes (enzymologie)</term>
<term>Séquence d'acides aminés</term>
<term>Séquence nucléotidique</term>
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<term>Isoenzymes</term>
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<term>Cathepsins</term>
<term>Endopeptidases</term>
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<term>Isoenzymes</term>
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<term>Base Sequence</term>
<term>Cathepsin B</term>
<term>Cysteine Endopeptidases</term>
<term>Humans</term>
<term>Nucleic Acid Hybridization</term>
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<term>Cysteine endopeptidases</term>
<term>Humains</term>
<term>Hybridation d'acides nucléiques</term>
<term>Séquence d'acides aminés</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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