Rat cathepsin H-catalyzed transacylation: Comparisons of the mechanism and the specificity with papain-superfamily proteases

Hironobu Koga, Nobuko Mori, Hidenori Yamada, Yukio Nishimura, Kazuo Tokuda, Keitaro Kato, Taiji Imoto

研究成果: ジャーナルへの寄稿記事

4 引用 (Scopus)

抄録

We found that rat cathepsin H showed strong transacylation activity under physiological conditions. It is a feature of cathepsin H to utilize amino acid amides not only as acylacceptors but also as acyl-donors in the reaction. The pH-dependence of the transacylation activity was distinct from those of other papain-superfamily proteases. The alkaline limb (pKapp=7.5) could be regarded as the pKa, of the α-amino group of the acyl-donor, which was also involved in the original amino-peptidase activity. The acidic limb (pKapp=5.8) was suggested to be involved in the deacylation step, where amino acid amide attacked the acyl-intermediate as a nucleophile in place of water in the hydrolysis. Although the Nα-deprotonated acyl-acceptor, which is supposed to govern the nucleophilic attack, has a small population in the acidic pH range (above pH 5), the transacylation was detectable even at the acidic pH-range because of the high S1' -site binding ability and suitable nucleophilicity of the acyl-acceptor. In the transacylation between various amino acid amides, the S1 and S1' site appeared to prefer hydrophobic residues without and regardless of a branch at β-carbon, respectively. From these results and the sequence homology in the papain superfamily, we concluded that the reaction was governed by the acyl-donor having a protonated amino group, the acyl-acceptor having a deprotonated amino group and the remarkable hydrophobic character (especially favoring tryptophan amide) of the S1' site, presumably reflecting the good conservation of Trpl77 in papain-superfamily proteases.

元の言語英語
ページ(範囲)939-944
ページ数6
ジャーナルJournal of Biochemistry
110
発行部数6
DOI
出版物ステータス出版済み - 1 1 1991

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Cathepsin H
Papain
Amides
Rats
Peptide Hydrolases
Amino Acids
Extremities
Nucleophiles
Sequence Homology
Tryptophan
Hydrolysis
Conservation
Carbon
Binding Sites
Water
Population

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Rat cathepsin H-catalyzed transacylation : Comparisons of the mechanism and the specificity with papain-superfamily proteases. / Koga, Hironobu; Mori, Nobuko; Yamada, Hidenori; Nishimura, Yukio; Tokuda, Kazuo; Kato, Keitaro; Imoto, Taiji.

:: Journal of Biochemistry, 巻 110, 番号 6, 01.01.1991, p. 939-944.

研究成果: ジャーナルへの寄稿記事

Koga, Hironobu ; Mori, Nobuko ; Yamada, Hidenori ; Nishimura, Yukio ; Tokuda, Kazuo ; Kato, Keitaro ; Imoto, Taiji. / Rat cathepsin H-catalyzed transacylation : Comparisons of the mechanism and the specificity with papain-superfamily proteases. :: Journal of Biochemistry. 1991 ; 巻 110, 番号 6. pp. 939-944.
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AU - Imoto, Taiji

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AB - We found that rat cathepsin H showed strong transacylation activity under physiological conditions. It is a feature of cathepsin H to utilize amino acid amides not only as acylacceptors but also as acyl-donors in the reaction. The pH-dependence of the transacylation activity was distinct from those of other papain-superfamily proteases. The alkaline limb (pKapp=7.5) could be regarded as the pKa, of the α-amino group of the acyl-donor, which was also involved in the original amino-peptidase activity. The acidic limb (pKapp=5.8) was suggested to be involved in the deacylation step, where amino acid amide attacked the acyl-intermediate as a nucleophile in place of water in the hydrolysis. Although the Nα-deprotonated acyl-acceptor, which is supposed to govern the nucleophilic attack, has a small population in the acidic pH range (above pH 5), the transacylation was detectable even at the acidic pH-range because of the high S1' -site binding ability and suitable nucleophilicity of the acyl-acceptor. In the transacylation between various amino acid amides, the S1 and S1' site appeared to prefer hydrophobic residues without and regardless of a branch at β-carbon, respectively. From these results and the sequence homology in the papain superfamily, we concluded that the reaction was governed by the acyl-donor having a protonated amino group, the acyl-acceptor having a deprotonated amino group and the remarkable hydrophobic character (especially favoring tryptophan amide) of the S1' site, presumably reflecting the good conservation of Trpl77 in papain-superfamily proteases.

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