Folding and Intramembraneous BRICHOS Binding of the Prosurfactant Protein C Transmembrane Segment

Abstract Surfactant protein C (SP-C) is a novel amyloid protein found in the lung tissue of patients suffering from interstitial lung disease (ILD) due to mutations in the gene of the precursor protein proSP-C. SP-C is a small α-helical hydrophobic protein with an unusually high content of valine residues. SP-C is prone to convert into β-sheet aggregates, forming amyloid fibrils. Nature's way of solving this folding problem is to include a BRICHOS domain in proSP-C, which functions as a chaperone for SP-C during biosynthesis. Mutations in the proSP-C BRICHOS domain or linker region lead to amyloid formation of the SP-C protein and ILD. In this study, we used an in vitro transcription/translation system to study translocon-mediated folding of the WT proSP-C poly-Val and a designed poly-Leu transmembrane (TM) segment in the endoplasmic reticulum (ER) membrane. Furthermore, to understand how the proSP-C BRICHOS domain present in the ER lumen can interact with the TM segment of proSP-C, we studied the membrane insertion properties of the recombinant form of proSP-C BRICHOS domain and two ILD-associated mutants. The results show that the co-translational folding of the WT proSP-C TM segment is inefficient, that the BRICHOS domain inserts into superficial parts of fluid membranes, and that BRICHOS membrane insertion is promoted by poly-Val peptides present in the membrane. In contrast, one BRICHOS and one non-BRICHOS ILD-associated mutant could not insert into membranes. These findings support a chaperone function of the BRICHOS domain, possibly together with the linker region, during proSP-C biosynthesis in the ER.