Abstract
Membrane recruitment of adaptor proteins is crucial for coupling antigen receptors to downstream signaling events. Despite the essential function of the B cell adaptor SLP-65, the mechanism of its recruitment to the plasma membrane is not yet understood. Here we show that a highly conserved leucine zipper in the SLP-65 N terminus is responsible for membrane association. Alterations in the N terminus abolished SLP-65 membrane localization and activity, both of which were restored by replacement of the N terminus with a myristoylation signal. The N terminus is an autonomous domain that confers specific localization and function when transferred to green fluorescent protein or the adaptor protein SLP-76. Our data elucidate the mechanism of SLP-65 membrane recruitment and suggest that leucine zipper motifs are essential interaction domains of signaling proteins.
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Acknowledgements
We thank C. Eschbach and I. Fidler for technical assistance. Supported by the Deutsche Forschungsgemeinschaft (SFB 620 and SFB 388).
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Supplementary information
Supplementary Fig. 1
The SLP-65-GFP and DNSLP-65-GFP fusion proteins were expressed comparably. (PDF 364 kb)
Supplementary Fig. 2
Membrane association of SLP-65 is independent of BCR expression. (PDF 948 kb)
Supplementary Fig. 3
Comparable expression of the different SLP-65 constructs in pre-B cells. (PDF 486 kb)
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Köhler, F., Storch, B., Kulathu, Y. et al. A leucine zipper in the N terminus confers membrane association to SLP-65. Nat Immunol 6, 204–210 (2005). https://doi.org/10.1038/ni1163
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DOI: https://doi.org/10.1038/ni1163
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