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.
Rosenkrands
I
I
Department of Biochemistry and Nutrition, Technical University of Denmark, Lyngby.
Hejgaard
J
J
Rasmussen
S K
SK
Bjørn
S E
SE
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D015843:Serpins / Q000302:isolation & purification* / Q000378:metabolism; D014908:Triticum / Q000737:chemistry*
D015843:Serpins