Fig. 1: HTRA1 interface mutants exhibit oligomeric assembly defects.
From: Rational correction of pathogenic conformational defects in HTRA1
a Schematic representation of HTRA1 domain organization and position of selected pathogenic mutations and the active site mutation S328A. Mutations located at the protomer-protomer interface are marked by an asterisk. The N-terminal (‘N’) domain consists of a fragmented IGFBP7 domain where neither the incomplete IGFBP binds insulin nor the incomplete Kazal-like domain functions as a protease inhibitor. b Position of pathogenic mutations in the HTRA1 trimer (PDB ID 3TJO). c Proteolytic activity of wt and mutant HTRA1s (1 µM) using β-casein (20 µM) as a substrate. The graph depicts the relative β-casein signal intensity (average ± SD of 3 independent experimental data). d Oligomeric states of wt and mutant HTRA1s evaluated by SEC-MALS. Upper panel: representative UV chromatograms of wt and mutant HTRA1s depicted in overlay. Lower panel: calculated molecular weights and relative abundance of mono-, di- and trimeric HTRA1 species. **: degradation products or impurities. e Oligomeric states of wt and mutant HTRA1s evaluated by native MS. Upper panels: representative spectra of HTRA1 wt and R274Q (5 µM). Mono- and trimeric species and charge states are labeled. Lower panel: relative abundance of mono- and trimeric HTRA1 species (5 µM; nd: not detected; mean + SD of 2 [wt, R166H] or 3 [R274Q, A173T, A252T] independent datasets; empty circles: individual data points). Dimers (relative abundance <5%) are not depicted. Source data are provided as a Source Data file.
