Abstract
Specific sorting signals direct transmembrane proteins to the compartments of the endosomal–lysosomal system1. Acidic-cluster-dileucine signals present within the cytoplasmic tails of sorting receptors, such as the cation-independent and cation-dependent mannose-6-phosphate receptors, are recognized by the GGA (Golgi-localized, γ-ear-containing, ADP-ribosylation-factor-binding) proteins2,3,4,5. The VHS (Vps27p, Hrs and STAM) domains6 of the GGA proteins are responsible for the highly specific recognition of these acidic-cluster-dileucine signals7,8,9,10. Here we report the structures of the VHS domain of human GGA3 complexed with signals from both mannose-6-phosphate receptors. The signals bind in an extended conformation to helices 6 and 8 of the VHS domain. The structures highlight an Asp residue separated by two residues from a dileucine sequence as critical recognition elements. The side chains of the Asp-X-X-Leu-Leu sequence interact with subsites consisting of one electropositive and two shallow hydrophobic pockets, respectively. The rigid spatial alignment of the three binding subsites leads to high specificity.
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Acknowledgements
We thank K. R. Rajashankar, U. Ramagopal, R. Trievel and E. Jones for assistance with data collection at beamline X9B, NSLS, D. E. Anderson for mass spectrometry, A. Peterkofsky for use of and assistance with the titration calorimeter, R. Ghirlando for analytical ultracentrifugation, X. Zhu for sequencing services, and H. Bernstein and P. Sun for comments on the manuscript.
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Misra, S., Puertollano, R., Kato, Y. et al. Structural basis for acidic-cluster-dileucine sorting-signal recognition by VHS domains. Nature 415, 933–937 (2002). https://doi.org/10.1038/415933a
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DOI: https://doi.org/10.1038/415933a
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