Limulus intracellular coagulation inhibitor type 3. Purification, characterization, cDNA cloning, and tissue localization

Kishan Lal Agarwala, Shun Ichiro Kawabata, Yoshiki Miura, Yuka Kuroki, Sadaaki Iwanaga

研究成果: ジャーナルへの寄稿記事

36 引用 (Scopus)

抄録

We reported that limulus intracellular coagulation inhibitor type-1 (LICI-1) (Miura, Y., Kawabata, S., and Iwanaga, S. (1994) J. Biol. Chem. 269, 542-547) and LICI type-2 (LICI-2) (Miura, Y., Kawabata, S., Wakamiya, Y., Nakamura, T., and Iwanaga, S. (1995) J. Biol. Chem. 270, 558-565) found in the hemocyte lysate belong to the serpin family. The LICI-1 specifically inhibits limulus lipopolysaccharide-sensitive serine protease, factor C (k1 = 2.5 x 106 M-1 s-1), whereas LICI-2 inhibits preferentially limulus clotting enzyme (k1 = 4.3 x 105 M-1 s-1). In our ongoing studies on limulus serpin, we found another inhibitor, named LICI type-3 (LICI-3), which strongly inhibits (1,3)-β-D-glucan-sensitive serine protease, factor G (k1 = 3.9 x 105 M-1 s-1). Thus, the limulus hemolymph coagulation cascade is effectively regulated by at least the three endogenous serpins. LICI-3, newly identified in hemocytes, is a single chain glycoprotein with an apparent M(r) = 53,000, the largest one among known limulus serpins. A cDNA sequence for LICI-3 coded a mature protein of 392 amino acids, of which 68 residues were confirmed by peptide sequencing. LICI-3 showed significant sequence similarity to LICI-1 (45.8% identity) and LICI-2 (33.7% identity). LICI-3 contained a putative reactive site, -Arg-Ser-, distinct from that of LICI-2 (-Lys-Ser-) but the same as that of LICI-1. Expression of LICI-3 mRNA was detected only in hemocytes, and not in heart, brain, stomach, intestine, coxal gland, and skeletal muscle. Immunoblotting of the hemocyte-derived large and small granules with antiserum against LICI-3 suggested that it is stored specifically in large granules, as in the case of LICI-1 and LICI-2, and is released in response to external stimuli.

元の言語英語
ページ(範囲)23768-23774
ページ数7
ジャーナルJournal of Biological Chemistry
271
発行部数39
DOI
出版物ステータス出版済み - 10 10 1996

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Horseshoe Crabs
Cloning
Coagulation
Serpins
Purification
Organism Cloning
Complementary DNA
Tissue
Hemocytes
Serine Proteases
Lipopolysaccharides
Muscle
Immune Sera
Brain
Glycoproteins
Hemolymph
Amino Acids
Messenger RNA
Peptides
Immunoblotting

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

これを引用

Limulus intracellular coagulation inhibitor type 3. Purification, characterization, cDNA cloning, and tissue localization. / Agarwala, Kishan Lal; Kawabata, Shun Ichiro; Miura, Yoshiki; Kuroki, Yuka; Iwanaga, Sadaaki.

:: Journal of Biological Chemistry, 巻 271, 番号 39, 10.10.1996, p. 23768-23774.

研究成果: ジャーナルへの寄稿記事

Agarwala, Kishan Lal ; Kawabata, Shun Ichiro ; Miura, Yoshiki ; Kuroki, Yuka ; Iwanaga, Sadaaki. / Limulus intracellular coagulation inhibitor type 3. Purification, characterization, cDNA cloning, and tissue localization. :: Journal of Biological Chemistry. 1996 ; 巻 271, 番号 39. pp. 23768-23774.
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abstract = "We reported that limulus intracellular coagulation inhibitor type-1 (LICI-1) (Miura, Y., Kawabata, S., and Iwanaga, S. (1994) J. Biol. Chem. 269, 542-547) and LICI type-2 (LICI-2) (Miura, Y., Kawabata, S., Wakamiya, Y., Nakamura, T., and Iwanaga, S. (1995) J. Biol. Chem. 270, 558-565) found in the hemocyte lysate belong to the serpin family. The LICI-1 specifically inhibits limulus lipopolysaccharide-sensitive serine protease, factor C (k1 = 2.5 x 106 M-1 s-1), whereas LICI-2 inhibits preferentially limulus clotting enzyme (k1 = 4.3 x 105 M-1 s-1). In our ongoing studies on limulus serpin, we found another inhibitor, named LICI type-3 (LICI-3), which strongly inhibits (1,3)-β-D-glucan-sensitive serine protease, factor G (k1 = 3.9 x 105 M-1 s-1). Thus, the limulus hemolymph coagulation cascade is effectively regulated by at least the three endogenous serpins. LICI-3, newly identified in hemocytes, is a single chain glycoprotein with an apparent M(r) = 53,000, the largest one among known limulus serpins. A cDNA sequence for LICI-3 coded a mature protein of 392 amino acids, of which 68 residues were confirmed by peptide sequencing. LICI-3 showed significant sequence similarity to LICI-1 (45.8{\%} identity) and LICI-2 (33.7{\%} identity). LICI-3 contained a putative reactive site, -Arg-Ser-, distinct from that of LICI-2 (-Lys-Ser-) but the same as that of LICI-1. Expression of LICI-3 mRNA was detected only in hemocytes, and not in heart, brain, stomach, intestine, coxal gland, and skeletal muscle. Immunoblotting of the hemocyte-derived large and small granules with antiserum against LICI-3 suggested that it is stored specifically in large granules, as in the case of LICI-1 and LICI-2, and is released in response to external stimuli.",
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