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Structural and functional characterization of human Iba proteins.
Schulze, Jörg O Quedenau, Claudia Roske, Yvette Adam, Thomas Schüler, Herwig Behlke, Joachim Turnbull, Andrew P Sievert, Volker Scheich, Christoph Mueller, Uwe
Schulze, Jörg O
Quedenau, Claudia
Roske, Yvette
Adam, Thomas
Schüler, Herwig
Behlke, Joachim
Turnbull, Andrew P
Sievert, Volker
Scheich, Christoph
Mueller, Uwe
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2008-09
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Abstract
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 Ca2+. 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 Ca2+. Furthermore, no binding of Ca2+ up to 0.1 mM was detected by equilibrium dialysis. Correspondingly, Iba EF-hand motifs lack residues essential for strong Ca2+ 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.
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Structural and functional characterization of human Iba proteins. 2008, 275 (18):4627-40 FEBS J.
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en
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1742-464X