Cloning and expression of the gene encoding Acacia confusa trypsin inhibitor that is active without post-translational proteolysis.
A recombinant plasmid containing the coding regions for Acacia confusa trypsin inhibitor (ACTI) has been constructed and expressed in Escherichia coli cells, as a fusion protein between ACTI and glutathione S-transferase (GST). The GST-fusion was produced as a soluble protein which did not require denaturing agents such as urea to solubilize it. The recombinant ACTI (reACTI) was obtained by treating the GST-fusion protein with thrombin. Both the reACTI and fusion protein have a strong inhibitory effect on trypsin activity without post-translational proteolysis.
Institute of Biochemistry, College of Medicine, National Taiwan University, Taipei, ROC.
C H
CH
Hung
M C
MC
Lee
M T
MT
Lin
J Y
JY
Lin
D000045:Acacia; D000595:Amino Acid Sequence; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D017343:Genes, Plant; D005982:Glutathione Transferase / Q000302:isolation & purification; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D010944:Plants / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000494:pharmacology; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D015183:Restriction Mapping; D014361:Trypsin Inhibitors / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology
D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; C074637:ACTI protein, Acacia confusa; D005982:Glutathione Transferase