Proposed lipocalin fold for apolipoprotein M based on bioinformatics and site-directed mutagenesis

Apolipoprotein RI (apoM) is a novel apolipoprotein that is predominantly present in high-density lipoprotein, Sensitive sequence starches, threading and comparative model building experiments revealed apoM to be structurally related to the lipocalin protein family. In a 3D model, characterized by an eight-stranded anti-parallel beta -barrel, a segment including Asn135 could adopt a closed or open conformation. Using site-directed mutagenesis, we demonstrated Asn135 in wild-type apoM to be glycosylated, suggesting that the segment is solvent exposed, ApoM displays two strong acidic patches of potential functional importance, one around the N-terminus and the other next to the opening of the beta -barrel. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B,V. All rights reserved.