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Metalloendopeptidase Meprin - Degradomic Approaches
Немски език
173 b
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In the past, singly-selected protease-substrates§(proteins cleaved by enzymes) were tested by in §vitro cleavage assays and thus substrate discovery §was haphazard. Nowadays, with the rapidly growing §field of proteomics (study of entire protein sub-§sets of a cell), a protease and its substrate §repertoire (degradome) may be studied in a complex §context to identify physiologically relevant §substrates. The author Daniel Ambort describes step-§by-step first the principles of degradomics and then §its application in protease-substrate discovery. Two §biological systems, a cell-based (culture medium-§derived proteins) and a cell-free (human milk as §polysubstrate), were used to find hitherto unkown §substrates for the metalloendopeptidase meprin (zinc-§dependent protein-cleaving enzyme). One- and two-§dimensional gel electrophoresis together with mass §spectrometry enabled the identification of novel §meprin cleavage products. In summary, this book §combines standard textbook knowledge with laboratory-§intense hard work and it thus encompasses a complete §description of the realization of degradomics §projects.
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