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. 2011 Dec 28;6(12):e29584. doi: 10.1371/journal.pone.0029584

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Pantakani et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Alignment of the FYVE domain of ZFYVE27 (ZFY27) with other FYVE family proteins; EEA1, ZFYVE9 (ZFYV9), HGS/HRS and yeast FAB1, PEP7, VPS27. The red boxes are highlighting the conserved WXXD motif (1), R(R/K)HHCR motif (2) and RVC motif (3). The conserved cysteine residues which co-ordinates the zinc ion binding were marked with asterisks. (B) The liposomal binding assay shows specific binding of c-Myc-ZFYVE27^WT to the PolyPIPosomes consisting of PtdIns3P. As a positive control, we also detected the binding of endogenous EEA1 to the PolyPIPosomes. (C) The recombinant GST-ZFYVE27^300-404 protein also showed binding affinity to PtdIns3P. As a negative control, GST (alone) protein was used in the liposomal binding assay.