Fig. 2: Metallo-IgA1P cleavage of IgA1.
From: A substrate-induced gating mechanism is conserved among Gram-positive IgA1 metalloproteases
a Proteolytic products of IgA1 after incubation with either G. haemolysans IgA1P WT or the E1848A mutant for the indicated time points comprised 20 μM IgA1 with 5 μM of each IgA1P. b Comparing the IgA1 cleavage pattern of G. haemolysans IgA1P with (residues 651-2201) or without (residues 907-2201) its N-terminal GhTrp domain. c Quantitative analysis of metallo-IgA1P cleavage activity of IgA1. Shown are the average of duplicate reads from an ELISA-based assay plated with the indicated construct, incubated for 30 min with IgA1, and quantified for remaining IgA1-FAB and normalized with the quantified bound IgA1-FC. These include S. pneumoniae (Sp) residues 665-1963 (black straight line), G. haemolysans (Gh) residues 907-2201 (red straight line), G. haemolysans residues 651-2201 (green straight line). d Quantitative analysis of IgA1 binding affinity to G. haemolysans IgA1P-E1848A and S. pneumoniae IgA1P-E1605A. Shown are the average of duplicate reads from an ELISA-based assay plated with 100 nM of each metallo-IgA1P mutant and incubated with the indicate concentration of IgA1.
