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A recombinant wheat serpin with inhibitory activity.
- Source :
-
Plant molecular biology [Plant Mol Biol] 1996 Feb; Vol. 30 (3), pp. 673-7. - Publication Year :
- 1996
-
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.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Carbohydrate Sequence
DNA, Plant
Escherichia coli
Humans
Molecular Sequence Data
Plant Proteins pharmacology
Recombinant Proteins genetics
Recombinant Proteins pharmacology
Serine Proteinase Inhibitors pharmacology
Serpins pharmacology
Plant Proteins genetics
Serine Proteinase Inhibitors genetics
Serpins genetics
Triticum genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0167-4412
- Volume :
- 30
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Plant molecular biology
- Publication Type :
- Academic Journal
- Accession number :
- 8605317
- Full Text :
- https://doi.org/10.1007/BF00049343