A Novel Influenza A Virus Activating Enzyme from Porcine Lung: Purification and Characterization
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Abstract
Proteolytic activation of hemagglutinin, an envelope glycoprotein of the influenza virus, by host proteases is essential for infection and proliferation of the virus. However, there is no well-defined, inherent source of host proteases in man or swine, both of which are natural hosts for human influenza viruses. We have recently isolated a 32 kDa protein in a high salt extract from porcine lungs, which possess the hemagglutinin processing activity. In this study, we attempted to purify another hemagglutinin processing enzyme from porcine lung. The purified enzyme, named tryptase TC30, exhibited a molecular mass of about 30 kDa by SDS-PAGE and 28.5 kDa by gel filtration chromatography, suggesting that it is a monomer. Tryptase TC30 cleaved peptide substrates with Arg at the P1 position, and preferentially substrates with the Ser-Ile-Gln-Ser-Arg sequence corresponding to the HA cleavage site sequence of the A/PR/8/34 influenza virus. Among various inhibitors tested, trypsintype serine protease inhibitors, such as aprotinin, antipain, benzamidine and leupeptin, efficiently inhibited the proteolytic activity of the enzyme. The N-terminal 40 amino acid sequence of tryptase TC30 exhibits more than 60% homology to mast cell tryptases from mice MCP-6 and human tryptase-α and -β. These data indicate that tryptase TC30, the 30 kDa enzyme from porcine lung, is a novel hemagglutinin-cleaving enzyme.
Copyright © 2003 by Walter de Gruyter GmbH & Co. KG
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Artikel in diesem Heft
- Obituary
- Thrombin Signaling in the Brain: The Role of Protease-Activated Receptors
- Transcriptional Repression of the Human p53 Gene by Hepatitis B Viral Core Protein (HBc) in Human Liver Cells
- Identification of Cytosolic Leucyl Aminopeptidase (EC 3.4.11.1) as the Major Cysteinylglycine-Hydrolysing Activity in Rat Liver
- A Novel Influenza A Virus Activating Enzyme from Porcine Lung: Purification and Characterization
- Binding of Urokinase Plasminogen Activator to gp130 via a Putative Urokinase-Binding Consensus Sequence
- Modifications of Glyceraldehyde-3-Phosphate Dehydrogenase Induced by Increasing Concentrations of Peroxynitrite: Early Recognition by 20S Proteasome
- Different Isoforms of the Non-Integrin Laminin Receptor Are Present in Mouse Brain and Bind PrP
- Cell-Type Specific Targeting and Gene Expression Using a Variant of Polyoma VP1 Virus-Like Particles
- Quantitation of Membrane Type Serine Protease 1 (MT-SP1) in Transformed and Normal Cells
- Interaction of Heat Shock Protein 70 Peptide with NK Cells Involves the NK Receptor CD94
- Cystatins C, E/M and F in Human Pleural Fluids of Patients with Neoplastic and Inflammatory Lung Disorders
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- Purification and Primary Structure Determination of Human Lysosomal Dipeptidase
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