Journal article
Serpins from wheat grain
Wheat serpin genes have been identified by Southern blot hybridization with three distinct barley protein Z probes. Immunoblot analysis with a monoclonal antibody towards barley protein Z confirmed expression of related M(r) approximately 40 kDa proteins in wheat grain. The wheat serpins were extracted under reducing conditions and separated from beta-amylase and other seed proteins by thiophilic adsorption and anion-exchange chromatography.
One molecular form possessing chymotrypsin inhibitory activity was isolated in a reactive site cleaved form on a chymotrypsin affinity column. N-terminal amino acid sequences of a CNBr fragment and of the C-terminal peptide from the cleaved inhibitor (M(r) 4574 +/- 4 Da) verified homology with barley protein Z and mammalian serpins.
The native inhibitory serpin was demonstrated to form an SDS-stable complex with alpha-chymotrypsin.
| Language: | English |
|---|---|
| Publisher: | Wiley |
| Year: | 1994 |
| Pages: | 75-80 |
| ISSN: | 18733468 and 00145793 |
| Types: | Journal article |
| DOI: | 10.1016/0014-5793(94)80610-1 |
Amino Acid Sequence Amino acid sequence Animals BSZ, barley serpin protein Z Barley protein Z Blotting, Southern Chromatography, Gel Chromatography, Ion Exchange Chymotrypsin inhibitor Complex formation DTT, Dithiothreitol Electrophoresis, Polyacrylamide Gel Humans Molecular Sequence Data PVDF, polyvinylidene difluoride RP-HPLC, reversed-phase high-performance liquid chromatography SSC, standard saline citrate Sequence Alignment Serpin Serpins T-gel, thiophilic gel TFA, trifluoroacetic acid TLCK, tosyl-lysine chloromethyl ketone Triticum Triticum aestivum L WSZ, protein Z-like wheat serpin WSZCI, protein Z-like wheat serpin with chymotrypsin inhibitor activity
