PMID: 9524068Apr 2, 1998Paper

Recombinant expression of human mast cell proteases chymase and tryptase

Biological Chemistry
Z M WangN M Schechter

Abstract

Expression of recombinant human chymase and tryptase was achieved in a baculovirus-insect cell system using a fusion protein construct. Recombinant baculovirus was produced with DNA coding for a NH2-ubiquitin-chymase-COOH or NH2-ubiquitin-tryptase-COOH fusion protein inserted immediately downstream of the signal sequence for the secreted envelope protein, glycoprotein 67. In each construct, the natural prepropeptide sequence of the protease was replaced by the amino acid sequence for the enterokinase cleavage site of trypsinogen. High Five insect cells infected with either of the modified baculovirus produced mg quantities of each fusion protein per liter of culture. Treatment of the chymase-fusion protein with enterokinase or the tryptase-fusion protein with enterokinase in the presence of a highly charged polysaccharide (dextran sulfate or heparin) produced enzymatically active proteases with properties of the native enzymes. A procedure for the purification of mg quantities of recombinant chymase from infected-cell medium is presented.

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Citations

Dec 31, 2002·Journal of Molecular and Cellular Cardiology·Ervin G ErdösPeter A Deddish
Nov 9, 2000·American Journal of Respiratory and Critical Care Medicine·M J CarrB J Undem
Jan 15, 2004·Biological Chemistry·Trevor SelwoodNorman M Schechter
Aug 21, 2008·Protein Expression and Purification·Li LiuJames E Strickler
Sep 21, 2007·Journal of Molecular Biology·Mekdes DebelaWolfram Bode
Jul 6, 2010·Biochemical Pharmacology·Jukka KervinenLawrence de Garavilla
Aug 19, 2014·Protein Expression and Purification·Eliot T SmithDavid A Johnson
Feb 15, 2001·The Journal of Allergy and Clinical Immunology·H K MinL B Schwartz
Nov 3, 2012·Hematology/oncology Clinics of North America·Stephen J Fuller
Mar 30, 2006·The Journal of Antibiotics·Takuro ShimataniIkutaro Saji
Sep 29, 2004·Journal of Enzyme Inhibition and Medicinal Chemistry·David AlagilleMarie-Claude Viaud-Massuard
Jun 11, 2002·The Journal of Biological Chemistry·Michael I PlotnickNorman M Schechter
Nov 6, 2007·The Journal of Biological Chemistry·Jukka KervinenJohn Spurlino
Aug 12, 1999·Archives of Biochemistry and Biophysics·M WalterN M Schechter
Aug 28, 1999·Bioorganic & Medicinal Chemistry Letters·W C GroutasJ Tu
May 17, 2001·Bioorganic & Medicinal Chemistry·U NeumannM Gütschow

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