4TKN

Structure of the SNX17 FERM domain bound to the second NPxF motif of KRIT1

Summary for 4TKN

• Entry DOI: 10.2210/pdb4tkn/pdb
• Descriptor: Sorting nexin-17, Krev interaction trapped protein 1 (3 entities in total)
• Functional Keywords: ferm domain, npxy motif, npxf motif, protein transport-signaling protein complex, protein transport/signaling protein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 6
• Total formula weight: 103179.43

Authors

Stiegler, A.L.,Zhang, R.,Liu, W.,Boggon, T.J. (deposition date: 2014-05-27, release date: 2014-07-30, Last modification date: 2024-10-23)

Primary citation

Stiegler, A.L.,Zhang, R.,Liu, W.,Boggon, T.J.
Structural Determinants for Binding of Sorting Nexin 17 (SNX17) to the Cytoplasmic Adaptor Protein Krev Interaction Trapped 1 (KRIT1).
J.Biol.Chem., 289:25362-25373, 2014

PubMed Abstract: Sorting nexin 17 (SNX17) is a member of the family of cytoplasmic sorting nexin adaptor proteins that regulate endosomal trafficking of cell surface proteins. SNX17 localizes to early endosomes where it directly binds NPX(Y/F) motifs in the cytoplasmic tails of its target receptors to mediate their rates of endocytic internalization, recycling, and/or degradation. SNX17 has also been implicated in mediating cell signaling and can interact with cytoplasmic proteins. KRIT1 (Krev interaction trapped 1), a cytoplasmic adaptor protein associated with cerebral cavernous malformations, has previously been shown to interact with SNX17. Here, we demonstrate that SNX17 indeed binds directly to KRIT1 and map the binding to the second Asn-Pro-Xaa-Tyr/Phe (NPX(Y/F)) motif in KRIT1. We further characterize the interaction as being mediated by the FERM domain of SNX17. We present the co-crystal structure of SNX17-FERM with the KRIT1-NPXF2 peptide to 3.0 Å resolution and demonstrate that the interaction is highly similar in structure and binding affinity to that between SNX17 and P-selectin. We verify the molecular details of the interaction by site-directed mutagenesis and pulldown assay and thereby confirm that the major binding site for SNX17 is confined to the NPXF2 motif in KRIT1. Taken together, our results verify a direct interaction between SNX17 and KRIT1 and classify KRIT1 as a SNX17 binding partner.

PubMed: 25059659
DOI: 10.1074/jbc.M114.584011

Experimental method

X-RAY DIFFRACTION (3 Å)