title: Cloning, expression and characterisation of murine procathepsin E.
abstract: The cDNA encoding murine procathepsin E was isolated and sequenced and recombinant enzyme was produced in Escherichia coli. The activity of the purified recombinant mouse cathepsin E was characterised quantitatively using two synthetic peptide substrates and naturally occurring inhibitors. The majority of the recombinant enzyme was present as a homodimer (Mr approximately 80) in which the two monomers were linked by an intermolecular disulfide bond. By analogy to previous studies with human cathepsin E, this is most likely a consequence of the presence of a unique cysteine residue near the N-terminus of the mature proteinase. The availability of (i) recombinant murine enzyme in reasonable quantities and (ii) a full-length cDNA now enables structural investigations and attempts to generate 'knock-out' mice deficient in this important aspartic proteinase to be undertaken.
authors.affiliation: School of Molecular and Medical Biosciences, University of Wales, Cardiff, UK.
authors.forename: P J
authors.initials: PJ
authors.lastname: Tatnell
authors.forename: W E
authors.initials: WE
authors.lastname: Lees
authors.forename: J
authors.initials: J
authors.lastname: Kay
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D020059:Cathepsin E; D002403:Cathepsins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D004792:Enzyme Precursors; D011994:Recombinant Proteins; D002403:Cathepsins; C067581:procathepsin E; D020059:Cathepsin E