Biochimica et Biophysica Acta vol:1387 issue:1-2 pages:317-24
The substrate and peptide bond specificities of a purified wheat gluten aspartic proteinase (GlAP) are studied. GlAP shows maximum gluten hydrolysing activity at pH 3.0. At this pH, especially the wheat high molecular weight glutenin subunits (HMW-GS) and to a lesser extent the low molecular weight glutenin subunits and gliadins are hydrolysed. GlAP has no obvious effect on albumins and globulins. In its action on oxidised insulin B-chain, GlAP forms eight peptides and has high specificity for peptide bonds located between amino acid residues with large hydrophobic side chains (Leu, Phe, Tyr) but the peptide bond Glu13-Ala14 is also hydrolysed. Although structurally quite similar to a barley aspartic proteinase, the peptide bond specificity of GlAP towards oxidised insulin B-chain resembles slightly more that of a cardoon aspartic proteinase, cardosin B. HMW-GS 7, purified from cultivar Galahad-77, is rapidly hydrolysed by GlAP. N-Terminal amino acid sequence data show that GlAP cleaves at least one Met-Ile peptide bond at the end of the N-terminal domain and two Val-Leu peptide bonds in the repetitive domain of HMW-GS 7.