Kakizawa T.,Kyoto Prefectural University of Medicine |
Sanjoh A.,Protein Wave Co. Minoo |
Kobayashi A.,Kyoto Prefectural University of Medicine |
Hattori Y.,Kyoto Prefectural University of Medicine |
And 2 more authors.
Bioorganic and Medicinal Chemistry | Year: 2011
A recombinant form of BACE1 (β-site amyloid precursor protein cleaving enzyme-1) corresponding to positions 46-454 of the extracellular domain of the original membrane enzyme was prepared. The recombinant BACE1 (rBACE1) had the kinetic parameters Km = 5.5 μM and kcat = 1719 s -1. Using several libraries of substrates containing unnatural amino acids, the specificity of rBACE1 was evaluated. LC/MS of digests derived from the libraries clarified that a dodecapeptide containing unnatural amino acids at P2 to P1′ was a superior cleavage sequence. © 2011 Elsevier Ltd. All rights reserved. Source