Evaluation of superior BACE1 cleavage sequences containing unnatural amino acids

Taeko Kakizawa; Akira Sanjoh; Akane Kobayashi; Yasunao Hattori; Kenta Teruya; Kenichi Akaji

doi:10.1016/j.bmc.2011.03.056

Evaluation of superior BACE1 cleavage sequences containing unnatural amino acids

Taeko Kakizawa, Akira Sanjoh, Akane Kobayashi, Yasunao Hattori, Kenta Teruya, Kenichi Akaji

Research output: Contribution to journal › Article › peer-review

7 Citations (Scopus)

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 = 5.5 μM and k_cat = 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.

Original language	English
Pages (from-to)	2785-2789
Number of pages	5
Journal	Bioorganic and Medicinal Chemistry
Volume	19
Issue number	9
DOIs	https://doi.org/10.1016/j.bmc.2011.03.056
Publication status	Published - 2011 May 1
Externally published	Yes

Keywords

Amyloid peptide
BACE1
Substrate specificity
Unnatural amino acid

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Pharmaceutical Science
Drug Discovery
Clinical Biochemistry
Organic Chemistry

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10.1016/j.bmc.2011.03.056

Cite this

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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 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.",

keywords = "Amyloid peptide, BACE1, Substrate specificity, Unnatural amino acid",

author = "Taeko Kakizawa and Akira Sanjoh and Akane Kobayashi and Yasunao Hattori and Kenta Teruya and Kenichi Akaji",

note = "Funding Information: This work was supported by a Grant-in-aid for Young Scientists (B, 22790119 ) from MEXT (The Ministry of Education, Culture, Sports, Science and Technology) to T.K. ",

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AU - Kakizawa, Taeko

AU - Sanjoh, Akira

AU - Kobayashi, Akane

AU - Hattori, Yasunao

AU - Teruya, Kenta

AU - Akaji, Kenichi

N1 - Funding Information: This work was supported by a Grant-in-aid for Young Scientists (B, 22790119 ) from MEXT (The Ministry of Education, Culture, Sports, Science and Technology) to T.K.

PY - 2011/5/1

Y1 - 2011/5/1

N2 - 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.

AB - 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.

KW - Amyloid peptide

KW - BACE1

KW - Substrate specificity

KW - Unnatural amino acid

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Evaluation of superior BACE1 cleavage sequences containing unnatural amino acids

Abstract

Keywords

ASJC Scopus subject areas

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