Fig. 3: Neprosin activity against molecules relevant for coeliac disease.
From: Molecular and in vivo studies of a glutamate-class prolyl-endopeptidase for coeliac disease therapy
a SDS-PAGE analysis of gliadin exposed to increasing concentrations of pepsin (left), neprosin (centre) or neprosin plus pepsin (right). Figure representative of two independent experiments. b Curves depicting gliadin cleavage as in (a) over time measured by turbidimetry. c Mass spectra of, top to bottom, the 33-mer peptide (3912 Da); the 33-mer peptide after incubation with 0.5 μM neprosin for 0 min, 20 min and overnight; neprosin alone; and the 33-mer peptide after overnight incubation with 10 μM pepsin, which leaves the peptide intact. d Gliadin zymogram depicting the activity of neprosin (left lane) and the mature enzyme resulting from pro-neprosin self-activation (right lane). e Same as (d) but showing gelatin zymography. f Sequence of the 33-mer and extent of the six overlapping HLA-DQ2.5-binding epitopes as highlighted by red double arrows7. Glutamines susceptible to deamidation by transglutaminase are shown in purple circles11. The peptide corresponds to segment L76–F108 of α-gliadin (UniProt ID P18573). g Cleavage of the 33-mer peptide by neprosin over time proceeds according to two pathways (top and bottom). For the distinct panels of this figure, relevant source data are provided as a Source Data file if adequate.
