PMID: 8605317Feb 1, 1996Paper

A recombinant wheat serpin with inhibitory activity

Plant Molecular Biology
S K RasmussenJ Hejgaard

Abstract

A full-length clone encoding the wheat (Triticum aestivum L.) serpin WSZ1 was isolated from a cDNA library based on mRNA from immature grain. The 398 amino acid sequence deduced from the cDNA was corroborated by sequencing CNBr peptides of WSZ1 purified from resting grain. WSZ1 belongs to the subfamily of protein Z-type serpins and the amino acid sequence is 70% identical with the barley serpins BSZ4 and BSZx and 27-33% identical with human serpins such as alpha 1-proteinase inhibitor, antithrombin III, and plasminogen activator inhibitor. The cDNA was subcloned in the pET3d expression vector, equipped with a histidine affinity tag at the N-terminus and expressed in Escherichia coli BL(21) DE3 pLysS. Recombinant WSZ1 from the soluble fraction was partially purified on Ni-NTA agarose and MonoQ columns and shown to form SDS-stable complexes with alpha-chymotrypsin. Southern blots and amino acid sequencing indicated that only few serpins are encoded by wheat, but at least three distinct genes are expressed in the grain. Cleavage experiments on a chymotrypsin column suggested a Gln-Gln reactive site bond not previously observed in inhibitory serpins.

References

Jan 1, 1990·Methods in Enzymology·F W StudierJ W Dubendorff
Dec 12, 1990·European Journal of Biochemistry·A BrandtJ Hejgaard
Jan 1, 1989·Carlsberg Research Communications·R Lundgard, B Svensson
Apr 18, 1994·FEBS Letters·I RosenkrandsS E Bjørn
May 16, 1994·FEBS Letters·A J SchulzeR A Engh
Aug 3, 1993·Biochemistry·P C HopkinsS R Stone
Feb 20, 1993·Biochimica Et Biophysica Acta·S K Rasmussen

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Citations

Dec 7, 2007·Functional & Integrative Genomics·Thomas H Roberts, Jørn Hejgaard
Oct 17, 1996·Biochimica Et Biophysica Acta·S K RasmussenI Svendsen
Jun 10, 2011·PloS One·Fernando Alvarez-AlfagemeGuy Smagghe
Oct 10, 2006·Journal of Molecular Biology·Dominique VercammenFrank Van Breusegem
Sep 30, 1996·FEBS Letters·S W DahlJ Hejgaard
Oct 11, 1996·The Journal of Biological Chemistry·S W DahlJ Hejgaard
Nov 29, 1996·The Journal of Biological Chemistry·P MelletJ G Bieth

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