Figure 5.

Negatively charged amino acids bracketing the shedding cleavage site interfere with the shedding of short ALCAM. RAW 264.7 cells expressing N-terminally Halo-tagged substitution mutants of short ALCAM were treated with or without LPS for 60 min, and cell extracts and culture supernatants were subjected to Western blotting (IB) with an anti-HaloTag antibody. The amino acid sequences of the stalk regions of WT and substitution mutants (DD and KK) of short ALCAM are shown. Arrowhead, shedding cleavage site of short ALCAM. The substituted amino acid residues are highlighted.