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Design of sensitive fluorogenic substrates for human cathepsin D
Author(s) -
Gulnik Sergei V.,
Suvorov Leonid I.,
Majer Pavel,
Collins Jack,
Kane Bradley P.,
Johnson Donald G.,
Erickson John W.
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00886-7
Subject(s) - valine , chemistry , cysteine , isoleucine , fluorescence , cathepsin d , methionine , biochemistry , cathepsin , stereochemistry , amino acid , enzyme , leucine , physics , quantum mechanics
Cathepsin D is a lysosomal aspartic proteinase that has been implicated in several pathological processes such as breast cancer and Alzheimer's disease. We designed and synthesized a number of quenched fluorogenic substrates with P2 variations in the series AcEE(EDANS)KPIXFFRLGK(DABCYL)E‐NH 2 , where X=cysteine, methylcysteine, ethylcysteine, tert ‐butylcysteine, carboxymethylcysteine, methionine, valine or isoleucine. Most of the fluorogenic substrates exhibited greater k cat / K m ratios than the best cathepsin D substrates described so far. Differences in kinetic constants, which were rationalized using structure‐based modeling, might make certain substrates useful for particular applications, such as active site titrations or initial velocity determination using a fluorescent plate reader.