LAPSE:2022.0086
Published Article
LAPSE:2022.0086
The Effect of a Peptide Substrate Containing an Unnatural Branched Amino Acid on Chymotrypsin Activity
Yuuki Yamawaki, Tomoki Yufu, Tamaki Kato
October 18, 2022
7-Amino-4-methylcoumarin (AMC) is a low molecular weight fluorescent probe that can be attached to a peptide to enable the detection of specific proteases, such as chymotrypsin, expressed in certain diseases. Because this detection depends on the specificity of the protease toward the peptidyl AMC, the development of specific substrates is required. To investigate the specificity of chymotrypsin, peptidyl AMC compounds incorporating four different amino acid residues were prepared by liquid-phase synthesis. Two unnatural amino acids, 2-amino-4-ethylhexanoic acid (AEH) and cyclohexylalanine (Cha), were used to investigate the substrate specificity as these amino acids have structures different from natural amino acids. AEH was synthesized using diethyl acetamidemalonate as a starting material. The substrate containing Cha had high hydrophobicity and showed a high reaction velocity with chymotrypsin. Although the AEH substrate with a branched side chain had high hydrophobicity, it showed a low reaction velocity. The substrate containing the aromatic amino acid phenylalanine was less hydrophobic than the Cha and AEH substrates, but chymotrypsin showed the highest specificity for this compound. These results demonstrated that the substrate specificity of chymotrypsin is not only affected by the hydrophobicity and aromaticity, but also by the structural expanse of amino acid residues in the substrate.
Keywords
7-amino-4-methylcoumarin, chymotrypsin, fluorescent probe, peptide, specificity, unnatural amino acid
Subject
Suggested Citation
Yamawaki Y, Yufu T, Kato T. The Effect of a Peptide Substrate Containing an Unnatural Branched Amino Acid on Chymotrypsin Activity. (2022). LAPSE:2022.0086
Author Affiliations
Yamawaki Y: Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu 808-0196, Fukuoka, Japan
Yufu T: Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu 808-0196, Fukuoka, Japan
Kato T: Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu 808-0196, Fukuoka, Japan
Journal Name
Processes
Volume
9
Issue
2
First Page
242
Year
2021
Publication Date
2021-01-28
Published Version
ISSN
2227-9717
Version Comments
Original Submission
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PII: pr9020242, Publication Type: Journal Article
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LAPSE:2022.0086
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doi:10.3390/pr9020242
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Oct 18, 2022
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CC BY 4.0
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[v1] (Original Submission)
Oct 18, 2022
 
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Oct 18, 2022
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Mina Naeini
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