The optimal activity of a pseudozymogen form of recombinant matriptase under the mildly acidic pH and low ionic strength conditions.
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抄録
Matriptase is a transmembrane serine protease that is strongly expressed in epithelial cells. The single-chain zymogen of matriptase is considered to have inherent activity, leading to its own activation (i.e. conversion to the disulphide-linked-two-chain form by cleavage after Thr-Lys-Gln-Ala-Arg614). Also, there is growing evidence that the activation of zymogen occurs at the cell surface and in relation to the acidification and lowering of ionic strength within cell-surface microenvironments. The present study aimed to provide evidence for the involvement of zymogen activity in its activation in physiologically relevant cellular contexts. For this purpose, the activity of a pseudozymogen form of recombinant matriptase (HL-matriptase zymogen) was examined using acetyl-l-Lys-l-Thr-l-Lys-l-Gln-l-Leu-l-Arg-4-methyl-coumaryl-7-amide as a substrate. HL-matriptase zymogen exhibited optimal activity toward the substrate pH approximately 6.0. The substrate hydrolysis at the pH value was hardly detected when NaCl was present at a concentration of 145 mM. In a buffer of pH 6.0 containing 5 mM NaCl, the activity of HL-matriptase zymogen was only approximately 30-times lower than that of the respective two-chain form. These findings suggest that the in vivo activation of matriptase zymogen occurs via a mechanism involving the zymogen activity.
収録刊行物
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- Journal of biochemistry
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Journal of biochemistry 147 (4), 485-492, 2010-04
Oxford University Press on behalf of the Japanese Biochemical Society
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キーワード
- Amino Acid Chloromethyl Ketones/metabolism
- Amino Acid Motifs
- Animals
- Enteropeptidase
- Enzyme Activation
- Enzyme Precursors/chemistry
- Enzyme Precursors/genetics
- Enzyme Precursors/metabolism
- Hydrogen-Ion Concentration
- Kinetics
- Muscle Hypotonia
- Oligopeptides/chemistry
- Oligopeptides/metabolism
- Osmolar Concentration
- Protein Structure, Tertiary
- Rats
- Recombinant Fusion Proteins/chemistry
- Recombinant Fusion Proteins/metabolism
- Serine Endopeptidases/chemistry
- Serine Endopeptidases/genetics
- Serine Endopeptidases/metabolism
- Substrate Specificity
詳細情報 詳細情報について
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- CRID
- 1050282677042023936
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- NII論文ID
- 10027874930
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- NII書誌ID
- AA00694073
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- ISSN
- 0021924X
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- HANDLE
- 2433/128777
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- NDL書誌ID
- 10645061
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- 本文言語コード
- en
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- 資料種別
- journal article
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- データソース種別
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- IRDB
- NDL
- CiNii Articles