Isolation and characterization of Staphylocoagulase chymotryptic fragment. Localization of the procoagulant- and prothrombin-binding domain of this protein

Shun-Ichiro Kawabata, T. Morita, T. Miyata, S. Iwanaga, H. Igarashi

研究成果: ジャーナルへの寄稿記事

28 引用 (Scopus)

抄録

Several strains of Staphylococcus aereus secrete a protein, staphylocoagulase, that binds stoichiometrically to human prothrombin, resulting in a coagulant complex designated staphylothrombin. In the present study, staphylocoagulase was digested with α-chymotrypsin and the resulting fragments were isolated by gel filtration. One fragment (M(r) 43,000) exhibited a high affinity for human prothrombin (K(d) = 1.7 x 10-9 M), which is comparable to the affinity observed using intact staphylocoagulase (K(d) = 4.6 x 10-10 M). A complex of the M(r) 43,000 fragment and prothrombin possessed both clotting and amidase activity essentially identical to that observed in a complex of intact staphylocoagulase and prothrombin. A second fragment (M(r) 30,000) exhibited weaker affinity for prothrombin (K(d) = 1.2 x 10-7 M). While clotting activity was not observed with a complex of this fragment and prothrombin, it nonetheless possessed a weak amidase activity. A third fragment (M(r) 20,000) was found to bind to prothrombin, but the resultant complex did not exhibit clotting or amidase activity. Amino-terminal sequence analyses of these staphylocoagulase fragments revealed that the M(r) 43,000 fragment constitutes the amino-terminal portion of staphylocoagulase and also contains the M(r) 30,000 and 20,000 fragments. Moreover, the amino-terminal sequence of the M(r) 20,000 fragment was identical to that observed for the M(r) 30,000 fragment. From these results, we conclude that the functional region of staphylocoagulase for binding and activation of human prothrombin is localized in the amino-terminal region of the intact bacterial protein.

元の言語英語
ページ(範囲)1427-1433
ページ数7
ジャーナルJournal of Biological Chemistry
261
発行部数3
出版物ステータス出版済み - 7 9 1986

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Coagulase
Prothrombin
Carrier Proteins
amidase
Proteins
Coagulants
Bacterial Proteins
Chymotrypsin
Staphylococcus
Gel Chromatography
Sequence Analysis
Gels
Chemical activation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Isolation and characterization of Staphylocoagulase chymotryptic fragment. Localization of the procoagulant- and prothrombin-binding domain of this protein. / Kawabata, Shun-Ichiro; Morita, T.; Miyata, T.; Iwanaga, S.; Igarashi, H.

:: Journal of Biological Chemistry, 巻 261, 番号 3, 09.07.1986, p. 1427-1433.

研究成果: ジャーナルへの寄稿記事

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title = "Isolation and characterization of Staphylocoagulase chymotryptic fragment. Localization of the procoagulant- and prothrombin-binding domain of this protein",
abstract = "Several strains of Staphylococcus aereus secrete a protein, staphylocoagulase, that binds stoichiometrically to human prothrombin, resulting in a coagulant complex designated staphylothrombin. In the present study, staphylocoagulase was digested with α-chymotrypsin and the resulting fragments were isolated by gel filtration. One fragment (M(r) 43,000) exhibited a high affinity for human prothrombin (K(d) = 1.7 x 10-9 M), which is comparable to the affinity observed using intact staphylocoagulase (K(d) = 4.6 x 10-10 M). A complex of the M(r) 43,000 fragment and prothrombin possessed both clotting and amidase activity essentially identical to that observed in a complex of intact staphylocoagulase and prothrombin. A second fragment (M(r) 30,000) exhibited weaker affinity for prothrombin (K(d) = 1.2 x 10-7 M). While clotting activity was not observed with a complex of this fragment and prothrombin, it nonetheless possessed a weak amidase activity. A third fragment (M(r) 20,000) was found to bind to prothrombin, but the resultant complex did not exhibit clotting or amidase activity. Amino-terminal sequence analyses of these staphylocoagulase fragments revealed that the M(r) 43,000 fragment constitutes the amino-terminal portion of staphylocoagulase and also contains the M(r) 30,000 and 20,000 fragments. Moreover, the amino-terminal sequence of the M(r) 20,000 fragment was identical to that observed for the M(r) 30,000 fragment. From these results, we conclude that the functional region of staphylocoagulase for binding and activation of human prothrombin is localized in the amino-terminal region of the intact bacterial protein.",
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T1 - Isolation and characterization of Staphylocoagulase chymotryptic fragment. Localization of the procoagulant- and prothrombin-binding domain of this protein

AU - Kawabata, Shun-Ichiro

AU - Morita, T.

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AU - Igarashi, H.

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N2 - Several strains of Staphylococcus aereus secrete a protein, staphylocoagulase, that binds stoichiometrically to human prothrombin, resulting in a coagulant complex designated staphylothrombin. In the present study, staphylocoagulase was digested with α-chymotrypsin and the resulting fragments were isolated by gel filtration. One fragment (M(r) 43,000) exhibited a high affinity for human prothrombin (K(d) = 1.7 x 10-9 M), which is comparable to the affinity observed using intact staphylocoagulase (K(d) = 4.6 x 10-10 M). A complex of the M(r) 43,000 fragment and prothrombin possessed both clotting and amidase activity essentially identical to that observed in a complex of intact staphylocoagulase and prothrombin. A second fragment (M(r) 30,000) exhibited weaker affinity for prothrombin (K(d) = 1.2 x 10-7 M). While clotting activity was not observed with a complex of this fragment and prothrombin, it nonetheless possessed a weak amidase activity. A third fragment (M(r) 20,000) was found to bind to prothrombin, but the resultant complex did not exhibit clotting or amidase activity. Amino-terminal sequence analyses of these staphylocoagulase fragments revealed that the M(r) 43,000 fragment constitutes the amino-terminal portion of staphylocoagulase and also contains the M(r) 30,000 and 20,000 fragments. Moreover, the amino-terminal sequence of the M(r) 20,000 fragment was identical to that observed for the M(r) 30,000 fragment. From these results, we conclude that the functional region of staphylocoagulase for binding and activation of human prothrombin is localized in the amino-terminal region of the intact bacterial protein.

AB - Several strains of Staphylococcus aereus secrete a protein, staphylocoagulase, that binds stoichiometrically to human prothrombin, resulting in a coagulant complex designated staphylothrombin. In the present study, staphylocoagulase was digested with α-chymotrypsin and the resulting fragments were isolated by gel filtration. One fragment (M(r) 43,000) exhibited a high affinity for human prothrombin (K(d) = 1.7 x 10-9 M), which is comparable to the affinity observed using intact staphylocoagulase (K(d) = 4.6 x 10-10 M). A complex of the M(r) 43,000 fragment and prothrombin possessed both clotting and amidase activity essentially identical to that observed in a complex of intact staphylocoagulase and prothrombin. A second fragment (M(r) 30,000) exhibited weaker affinity for prothrombin (K(d) = 1.2 x 10-7 M). While clotting activity was not observed with a complex of this fragment and prothrombin, it nonetheless possessed a weak amidase activity. A third fragment (M(r) 20,000) was found to bind to prothrombin, but the resultant complex did not exhibit clotting or amidase activity. Amino-terminal sequence analyses of these staphylocoagulase fragments revealed that the M(r) 43,000 fragment constitutes the amino-terminal portion of staphylocoagulase and also contains the M(r) 30,000 and 20,000 fragments. Moreover, the amino-terminal sequence of the M(r) 20,000 fragment was identical to that observed for the M(r) 30,000 fragment. From these results, we conclude that the functional region of staphylocoagulase for binding and activation of human prothrombin is localized in the amino-terminal region of the intact bacterial protein.

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