Characterization of an aldolase-binding site in the Wiskott-Aldrich syndrome protein.
The thrombospondin-related anonymous protein (TRAP) is an essential transmembrane molecule in Plasmodium sporozoites. TRAP displays adhesive motifs on the extracellular portion, whereas its cytoplasmic tail connects to actin via aldolase, thus driving parasite motility and host cell invasion. The minimal requirements for the TRAP binding to aldolase were scanned here and found to be shared by different human proteins, including the Wiskott-Aldrich syndrome protein (WASp) family members. In vitro and in vivo binding of WASp members to aldolase was characterized by biochemical, deletion mapping, mutagenesis, and co-immunoprecipitation studies. As in the case of TRAP, the binding of WASp to aldolase is competitively inhibited by the enzyme substrate/products. Furthermore, TRAP and WASp, but not other unrelated aldolase binders, compete for the binding to the enzyme in vitro. Together, our results define a conserved aldolase binding motif in the WASp family members and suggest that aldolase modulates the motility and actin dynamics of mammalian cells. These findings along with the presence of similar aldolase binding motifs in additional human proteins, some of which indeed interact with aldolase in pull-down assays, suggest supplementary, non-glycolytic roles for this enzyme.
Buscaglia
Carlos A
CA
Michael Heidelberg Division of Immunology, Department of Pathology, New York University School of Medicine, New York, New York 10016, USA. buscac01@med.nyu.edu
Penesetti
Deepak
D
Tao
Mingyuan
M
Nussenzweig
Victor
V
D000135:Acid Phosphatase / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000138:chemical synthesis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D000071681:Tartrate-Resistant Acid Phosphatase; D051316:Wiskott-Aldrich Syndrome Protein / Q000235:genetics* / Q000378:metabolism*
D018076:DNA, Complementary; D007527:Isoenzymes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D051316:Wiskott-Aldrich Syndrome Protein; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D005634:Fructose-Bisphosphate Aldolase