The Brichos domain of prosurfactant protein C can hold and fold a transmembrane segment

Prosurfactant protein C (proSP‐C) is a 197‐residue integral membrane protein, in which the C‐terminal domain (CTC, positions 59–197) is localized in the endoplasmic reticulum (ER) lumen and contains a Brichos domain (positions 94–197). Mature SP‐C corresponds largely to the transmembrane (TM) region of proSP‐C. CTC binds to SP‐C, provided that it is in nonhelical conformation, and can prevent formation of intracellular amyloid‐like inclusions of proSP‐C that harbor mutations linked to interstitial lung disease (ILD). Herein it is shown that expression of proSP‐C (1–58), that is, the N‐terminal propeptide and the TM region, in HEK293 cells results in virtually no detectable protein, while coexpression of CTC in trans yields SDS‐soluble monomeric proSP‐C (1–58). Recombinant human (rh) CTC binds to cellulose‐bound peptides derived from the nonpolar TM region, but not the polar cytosolic part, of proSP‐C, and requires ≥5‐residues for maximal binding. Binding of rhCTC to a nonhelical peptide derived from SP‐C results in α‐helix formation provided that it contains a long TM segment. Finally, rhCTC and rhCTC Brichos domain shows very similar substrate specificities, but rhCTC^L188Q, a mutation linked to ILD is unable to bind all peptides analyzed. These data indicate that the Brichos domain of proSP‐C is a chaperone that induces α‐helix formation of an aggregation‐prone TM region.