Structural basis for dimerization of the Dictyostelium gelation factor (ABP120) rod

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Abstract

Gelation factor (ABP120) is one of the principal actin-cross-linking proteins of Dictyostelium discoideum. The extended molecule has an N-terminal 250-residue actin-binding domain and a rod constructed from six 100-residue repeats that have an Ig fold. The ability to dimerize is crucial to the actin cross-linking function of gelation factor and is mediated by the rod in which the two chains are arranged in an antiparallel fashion. We report the 2.2 Å resolution crystal structure of rod domains 5 and 6, which shows that dimerization is mediated primarily by rod domain 6 and is the result of a double edge-to-edge extension of β-sheets. Thus, contrary to earlier proposals, the chains of the dimeric gelation factor molecule overlap only within domain 6, and domains 1–5 do not pair with domains from the other chain. This information allows construction of a model of the gelation factor molecule and suggests how the chains in the related molecule filamin (ABP280) may interact.

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Figure 1: Structure of gelation factor.
Figure 2: Topology of gelation factor.
Figure 3: Sigma-A weighted 2Fo - Fc electron density maps.
Figure 4: Sequence alignments of gelation factor and ABP280.
Figure 5: Model of gelation factor.

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Acknowledgements

We thank A.Thompson, G. Leonard and A. Baker for assistance in collecting MAD data at BL14 at ESRF, Grenoble; R. Muller and H. Kent for assistance with protein preparation and light scattering; A. Murzin, E. de la Fortelle and J. Irwin for advice; and our colleagues in Martinsried and Cambridge, especially M. Schleicher, for their assistance. A.J.M. is a Howard Florey Fellow of the Royal Society of London. P.F. held a short-term EMBO Fellowship, and this work was supported in part by EC CHRX-CT93-0250 and a grant to A.N. from the Centre for Molecular Medicine, Cologne.

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Correspondence to Murray Stewart.

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