Processing of the precursors to neurotensin and other bioactive peptides by cathepsin E.
Cathepsin E (EC 3.4.23.34), an intracellular aspartic proteinase, was purified from monkey intestine by simple procedures that included affinity chromatography and fast protein liquid chromatography. Cathepsin E was very active at weakly acidic pH in the processing of chemically synthesized precursors such as the precursor to neurotensin/neuromedin, proopiomelanocortin, the precursor to xenopsin, and angiotensinogen. The processing sites were adjacent to a dibasic motif in the former two precursors and at hydrophobic recognition sites in the latter two. The common structural features that specified the processing sites were found in the carboxyl-terminal sequences of the active peptide moieties of these precursors; namely, the sequence Pro-Xaa-X'aa-hydrophobic amino acid was found at positions P4 through P1. Pro at the P4 position is thought to be important for directing the processing sites of the various precursor molecules to the active site of cathepsin E. Although the positions of Xaa and X'aa were occupied by various amino acids, including hydrophobic and aromatic amino acids, some of these had a negative effect, as typically observed when Glu/Arg and Pro were present at the P3 and P2 positions, respectively. Cathepsin D was much less active or was almost inactive in the processing of the precursors to neurotensin and related peptides as a result of the inability of the Pro-directed conformation of the precursor molecules to gain access to the active site of cathepsin D. Thus, the consensus sequence of precursors, Pro-Xaa-X'aa-hydrophobic amino acid, might not only generate the best conformation for cleavage by cathepsin E but might be responsible for the difference in specificities between cathepsins E and D.
Kageyama
T
T
Department of Cellular and Molecular Biology, Kyoto University, Aichi, Japan.
Ichinose
M
M
Yonezawa
S
S
D000595:Amino Acid Sequence; D000818:Animals; D020059:Cathepsin E; D002403:Cathepsins / Q000494:pharmacology*; D008251:Macaca; D008969:Molecular Sequence Data; D009496:Neurotensin / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D010455:Peptides; D011498:Protein Precursors / Q000378:metabolism*; D029867:Xenopus Proteins
D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D011498:Protein Precursors; D029867:Xenopus Proteins; C042090:neuromedin N; D009496:Neurotensin; C016832:xenopsin; D002403:Cathepsins; D020059:Cathepsin E