Serine proteinase inhibitor 3 and murinoglobulin I are potent inhibitors of neuropsin in adult mouse brain
| Autor: | Koji Takio, Atsunori Machida, Fiona L. Scott, Masao Shibata, Tomohiro Kamachi, Tatsuya Tanaka, Phillip I. Bird, Morito Akisada, Takuya Oka, Yoshimasa Miyake, Tadaaki Kishi, Jiuru Sun, Keiko Kato, Ryosuke Midorikawa, Kazuhiko Yamamoto, Kazumasa Matsumoto, Sadao Shiosaka, Naoshi Dohmae |
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| Předmět: |
Serine Proteinase Inhibitors
In situ hybridization Hippocampal formation Biology Biochemistry Hippocampus law.invention Serine Mice law Animals Molecular Biology Serine protease Cerebral Cortex Messenger RNA Hydrolysis Serine Endopeptidases Cell Biology Kallikrein Molecular biology Recombinant DNA biology.protein Kallikreins Serum Globulins |
| Zdroj: | Monash University |
| Popis: | Extracellular serine protease neuropsin (NP) is expressed in the forebrain limbic area of adult brain and is implicated in synaptic plasticity. We screened for endogenous NP inhibitors with recombinant NP (r-NP) from extracts of the hippocampus and the cerebral cortex in adult mouse brain. Two SDS-stable complexes were detected, and after their purification, peptide sequences were determined by amino acid sequencing and mass spectrometry, revealing that target molecules were serine proteinase inhibitor-3 (SPI3) and murinoglobulin I (MUG I). The addition of the recombinant SPI3 to r-NP resulted in an SDS-stable complex, and the complex formation followed bimolecular kinetics with an association rate constant of 3.4 +/- 0.22 x 10(6) M(-1) s(-1), showing that SPI3 was a slow, tight binding inhibitor of NP. In situ hybridization histochemistry showed that SPI3 mRNA was expressed in pyramidal neurons in the hippocampal CA1-CA3 subfields, as was NP mRNA. Alternatively, the addition of purified plasma MUG I to r-NP resulted in an SDS-stable complex, and MUG I inhibited degradation of fibronectin by r-NP to 24% at a r-NP/MUG I molar ratio of 1:2. Immunofluorescence histochemistry showed that MUG I localized in the hippocampal neurons. These findings indicate that SPI3 and MUG I serve to inactivate NP and control the level of NP in adult brain, respectively. |
| Databáze: | OpenAIRE |
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