Fig. 4.

Identification of novel O-glycosylated sialylated Thr residues in human adiponectin. A, FLAG-tagged headless (Stalk) and globular head (Glob) truncated proteins were either untreated or treated with neuraminidase before 2DE and Western blotting for the FLAG tag. B, 2DE of Thr to Ala mutants of human adiponectin secreted from transfected CHO cells. C, SDS-PAGE and Western blotting of the same samples. D, Velocity sedimentation analysis of secreted WT and T20-22A adiponectin. E, Sequence alignment of the adiponectin variable domain from various mammals is shown and reveals limited conservation of the modified sequence/residues (shaded). Data are representative of two to three independent experiments. H, Hexamers; T, trimers.