Cloning of a serine proteinase inhibitor from bovine brain: Expression in the brain and characterization of its target proteinases

Naoki Nakaya, Masahiro Nishibori, Masahiro Kawabata, Kiyomi Saeki

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

A cDNA encoding of the serine proteinase inhibitor (serpin), B-43, was cloned from the cDNA library of the bovine brain. It encoded 378 amino acids, and the MW of the protein was estimated to be 42.6 kDa, which is consistent with that of the native B-43 purified from the bovine brain. The homology search revealed that B-43 belongs to the ovalbumin branch of the serpin superfamily. Among them, B-43 was most homologous to human placental thrombin inhibitor (PI-6) and its murine counterpart, with the amino acid identity of 76% and 71%, respectively. Northern blot analysis showed that the size of the transcript was 1.4 kb, and that the expression of B-43 in the bovine brain varied depending on the brain regions, i.e. a lower level of expression was observed in the cerebral cortex and the hippocampus compared to the level of expression that was observed in the medulla oblongata. [35S]-labeled B-43 protein was synthesized in vitro by using a rabbit reticulocyte lysate system, which formed complexes with proteinases such as thrombin, trypsin, α-chymotrypsin, and 7S nerve growth factor (NGF), but not with urokinase or plasmin. These results, together with the immunohistochemical localization of B-43 in astrocytes and in some neurons which was observed in the previous study suggest that B-43 may be involved in the regulation of serine proteinases present in the brain or extravasated from the blood.

Original languageEnglish
Pages (from-to)293-300
Number of pages8
JournalMolecular Brain Research
Volume42
Issue number2
DOIs
Publication statusPublished - Dec 1996

Fingerprint

Serine Proteinase Inhibitors
Organism Cloning
Peptide Hydrolases
Brain
yeast proteinase B
Amino Acids
Medulla Oblongata
B 43
Fibrinolysin
Reticulocytes
Urokinase-Type Plasminogen Activator
Ovalbumin
Serine Proteases
Nerve Growth Factor
Gene Library
Thrombin
Astrocytes
Northern Blotting
Cerebral Cortex
Hippocampus

Keywords

  • B-43
  • Bovine brain
  • Cloning
  • Serine proteinase inhibitor
  • Thrombin

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Cloning of a serine proteinase inhibitor from bovine brain : Expression in the brain and characterization of its target proteinases. / Nakaya, Naoki; Nishibori, Masahiro; Kawabata, Masahiro; Saeki, Kiyomi.

In: Molecular Brain Research, Vol. 42, No. 2, 12.1996, p. 293-300.

Research output: Contribution to journalArticle

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