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Structural and functional characterization of human Iba proteins
Author(s) -
Schulze Jörg O.,
Quedenau Claudia,
Roske Yvette,
Adam Thomas,
Schüler Herwig,
Behlke Joachim,
Turnbull Andrew P.,
Sievert Volker,
Scheich Christoph,
Mueller Uwe,
Heinemann Udo,
Büssow Konrad
Publication year - 2008
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2008.06605.x
Subject(s) - actin , cytoskeleton , microbiology and biotechnology , actin binding protein , anthrax toxin , plasma protein binding , hela , chemistry , biophysics , fusion protein , biology , actin cytoskeleton , biochemistry , cell , recombinant dna , gene
Iba2 is a homolog of ionized calcium‐binding adapter molecule 1 (Iba1), a 17‐kDa protein that binds and cross‐links filamentous actin (F‐actin) and localizes to membrane ruffles and phagocytic cups. Here, we present the crystal structure of human Iba2 and its homodimerization properties, F‐actin cross‐linking activity, cellular localization and recruitment upon bacterial invasion in comparison with Iba1. The Iba2 structure comprises two central EF‐hand motifs lacking bound Ca 2+ . Iba2 crystallized as a homodimer stabilized by a disulfide bridge and zinc ions. Analytical ultracentrifugation revealed a different mode of dimerization under reducing conditions that was independent of Ca 2+ . Furthermore, no binding of Ca 2+ up to 0.1 m m was detected by equilibrium dialysis. Correspondingly, Iba EF‐hand motifs lack residues essential for strong Ca 2+ coordination. Sedimentation experiments and microscopy detected pronounced, indistinguishable F‐actin binding and cross‐linking activity of Iba1 and Iba2 with induction of F‐actin bundles. Fluorescent Iba fusion proteins were expressed in HeLa cells and co‐localized with F‐actin. Iba1 was recruited into cellular projections to a larger extent than Iba2. Additionally, we studied Iba recruitment in a Shigella invasion model that induces cytoskeletal rearrangements. Both proteins were recruited into the bacterial invasion zone and Iba1 was again concentrated slightly higher in the cellular extensions.