The structural basis of PTEN regulation by multi-site phosphorylation
Data files
Jun 27, 2022 version files 1.19 GB
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15N__13C__2H-ciVSP-C363S_SDS-PAGE_Gel.tif
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15N_VSP_C363S_Asn_unlabled_Gln_unlabled_R13G_SDS-PAGE_Gel.tif
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15N_VSP_C363S_His_unlabled_H122G_R158A_SDS-PAGE_Gel.tif
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15N_VSP_C363S_Lys_Arg_unlabled_R13G_SDS-PAGE_Gel.tif
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15N-VSP-C363S_SDS-PAGE_Gel.tif
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15N-VSP-C363S-Cys-tail-ligated_SDS-PAGE_Gel.tif
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15NVSP_Lys_unlabeled_R280G_K314A_SDS-PAGE_Gel.tif
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16mer-15N-tPTEN_Raw.raw
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1P_NMR_CSPs.xlsx
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3P_NMR_CSPs.xlsx
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4P_Intramol_NMR_CSPs.xlsx
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4P_NMR_CSPs.xlsx
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4p-crPTEN-22sp-T3_Raw.raw
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4p-VSP-C363S-T3_Raw.raw
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4pPTEN_13aalinker_15mer_SDS-PAGE_Gel.tif
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4pPTEN_16aalinker_18mer_crystal_SDS-PAGE_Gel.tif
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4pPTEN_20aalinker_18mer_tPTEN_13aalinker_SDS-PAGE_Gel.tif
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4pPTEN_crystal_22aa_spacer_18mer_tail_SDS-PAGE_Gel.tif
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Amino_Acid_Sequence_of_Key_Proteins.docx
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AP_assay_4p-PTEN_2.tif
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AP_assay_4p-PTEN_3.tif
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AP_assay_4p-PTEN.tif
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AP_assay_4p-VSP_1.tif
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AP_assay_4p-VSP_2.tif
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AP_assay_4p-VSP_3.tif
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cluster1_1.pdb
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cluster1_2.pdb
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cluster1_3.pdb
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cluster1_4.pdb
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cluster2_1.pdb
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cluster2_2.pdb
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cluster2_3.pdb
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cluster2_4.pdb
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DNA_Sequence_of_Key_Genes.docx
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N-Cys_PTEN_ND16_N-term_ligation_NMR_SDS-PAGE_Gel.tif
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nPTEN_13aalinker_15mer_SDS-PAGE_Gel.tif
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Peptide_resource_sheet.xlsx
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PIP2_NMR_CSPs.xlsx
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PRE_TEMPO_NMR.xlsx
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PTEN_Nterm_CSPs.xlsx
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PTEN_VSP_Binding_Assays_Raw_Data.xlsx
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PTEN_VSP_Enzyme_Assays_Raw_Data.xlsx
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README.xlsx
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tPTEN_N-term_mutants_WT_S10R_S10P_R15I_D24V_SDS-PAGE_Gel.tif
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tPTEN_Raw.raw
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tPTEN_SDS-PAGE_Gel.tif
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VSP-C363S_SDS-PAGE_Gel.tif
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VSP-C363S_VSP-C363S-K1364D-K367D_SDS-PAGE_Gel.tif
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VSP-Mxe_Chitin_4p-ligation_Concentration_gel_SDS-PAGE_Gel.tif
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Abstract
PTEN is a phosphatidylinositol-3,4,5-triphosphate (PIP3) phospholipid phosphatase that is commonly mutated or silenced in cancer. PTEN's catalytic activity, cellular membrane localization, and stability are orchestrated by a cluster of C-terminal phosphorylation events on Ser380, Thr382, Thr383, and Ser385, but the molecular details of this multifaceted regulation have remained uncertain. Here we use a combination of protein semisynthesis, biochemical analysis, NMR, X-ray crystallography, and computational simulations on human PTEN and its sea squirt homolog VSP to obtain a detailed picture of how the phospho-C-tail belts around PTEN's C2 and phosphatase domains. We also visualize a previously proposed dynamic N-terminal alpha-helix and show it is key for PTEN catalysis but disordered upon phospho-C-tail interaction. This structural model provides a comprehensive framework for how C-tail phosphorylation can impact PTEN's cellular functions.