title: Evaluation of superior BACE1 cleavage sequences containing unnatural amino acids.
abstract: 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 K(m)=5.5μM and k(cat)=1719s(-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 P(2) to [Formula: see text] was a superior cleavage sequence.
authors.affiliation: Department of Medicinal Chemistry, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kita-ku, Kyoto 603-8334, Japan.
authors.forename: Taeko
authors.initials: T
authors.lastname: Kakizawa
authors.forename: Akira
authors.initials: A
authors.lastname: Sanjoh
authors.forename: Akane
authors.initials: A
authors.lastname: Kobayashi
authors.forename: Yasunao
authors.initials: Y
authors.lastname: Hattori
authors.forename: Kenta
authors.initials: K
authors.lastname: Teruya
authors.forename: Kenichi
authors.initials: K
authors.lastname: Akaji
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D053829:Amyloid Precursor Protein Secretases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003546:Cysteine Endopeptidases / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000596:Amino Acids; D010455:Peptides; D011994:Recombinant Proteins; D053829:Amyloid Precursor Protein Secretases; D003546:Cysteine Endopeptidases; C019913:clostripain; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human