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Reconstitution of BNIP3/NIX-mitophagy initiation reveals hierarchical flexibility of the autophagy machinery

Adriaenssens E, Schaar S, Cook ASI, Stuke JFM, Sawa-Makarska J, Nguyen TN, Ren X, Schuschnig M, Romanov J, Khuu G, Lazarou M, Hummer G, Hurley JH, Martens S
bioRxiv 2024.08.28.609967 (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
223723pETDuet1_His-TEV-EGFPExpression of recombinant protein for purification
223724pETDuet1_6xHis-TEV-EGFP-FIP200-CTR(1429-1591aa)Expression of recombinant protein for purification
223725pETDuet1_6xHis-TEV-mCherry-WIPI2dExpression of recombinant protein for purification
223726pGEX-4T1_GST-LC3AExpression of recombinant protein for purification
223727pGEX-4T1_GST-LC3CExpression of recombinant protein for purification
223728pGEX-4T1_GST-GBRPExpression of recombinant protein for purification
223729pGEX-4T1_GST-GBRPL1Expression of recombinant protein for purification
223730pGEX-4T1_GST-GBRPL2Expression of recombinant protein for purification
223731pGenLenti V5-NIX Stable expression of protein in mammalian cell culture
223732pGenLenti V5-BNIP3 Stable expression of protein in mammalian cell culture
223733pETDuet1_NIX(1-182aa)-THROMBIN-GST (WT)Expression of recombinant protein for purification
223734pETDuet1_FUNDC1(1-50aa)-THROMBIN-GST (WT)Expression of recombinant protein for purification
223735pCAG_mCherry-ATG13 (WT)Expression of recombinant protein for purification
223736pETDuet1_NIX(1-182aa)-EGFP-TEV-GST (WT)Expression of recombinant protein for purification
223737pETDuet1_FUNDC1(1-50aa)-EGFP-TEV-GST (WT)Expression of recombinant protein for purification
223738pETDuet1_NIX(1-182aa)-THROMBIN-GST (W36A/L39A; deltaLIR)Expression of recombinant protein for purification
223739pETDuet1_FUNDC1(1-50aa)-THROMBIN-GST (Y18A/L21A; deltaLIR)Expression of recombinant protein for purification
223740pBIG1a_GST-TEV-CK2alpha_CK2betaExpression of recombinant protein for purification
223741pBIG1a_GST-TEV-CK2alpha'_CK2betaExpression of recombinant protein for purification
223742pLIB_GST-TEV-SRC(WT)Expression of recombinant protein for purification, codon optimized for Insect cells
223743pLIB_GST-TEV-SRC (Y530F)Expression of recombinant protein for purification
223744pETDuet1_BCL2L13(1-465aa)-THROMBIN-GSTExpression of recombinant protein for purification
223745pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GSTExpression of recombinant protein for purification
223746pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (W276A/I279A; deltaLIR1)Expression of recombinant protein for purification
223747pETDuet1_6xHis-TEV-Lambda-Protein-PhosphataseExpression of recombinant protein for purification
223748pETDuet1_NIX(1-182aa)-EGFP-TEV-GST (W36A/L39A; deltaLIR)Expression of recombinant protein for purification
223749pETDuet1_BCL2L13(1-465aa)-THROMBIN-GST (W276A/I279A; deltaLIR1)Expression of recombinant protein for purification
223750pETDuet1_FUNDC1(1-50aa)-EGFP-TEV-GST (Y18A/L21A; deltaLIR)Expression of recombinant protein for purification
223751pETDuet1_6xHis-TEV-mCherry-WIPI2d (K87A/K88A)Expression of recombinant protein for purification
223752pETDuet1_BCL2L13(1-465aa)-THROMBIN-GST (Y213A/I216A/W276A/I279A; deltaLIR1+2)Expression of recombinant protein for purification
223753pETDuet1_NIX(1-182aa)-THROMBIN-GST (E72A/L75A/D77A/E81A; deltaWIPI2)Expression of recombinant protein for purification
223754pETDuet1_BCL2L13(1-465aa)-THROMBIN-GST (I224A/L227A/W276A/I279A; deltaLIR1+3)Expression of recombinant protein for purification
223755pETDuet1_BCL2L13(1-465aa)-THROMBIN-GST (W276A/I279A/I307A/V310A; deltaLIR1+4)Expression of recombinant protein for purification
223756pETDuet1_BCL2L13(1-465aa)-THROMBIN-GST (I224A/L227A/W276A/I279A/I307A/V310A; deltaLIR1+3+4)Expression of recombinant protein for purification
223757pHAGE-FKBP-mEGFP-WIPI2Stable expression and inducible recruitment of protein in mammalian cell culture
223758pHAGE-FKBP-mEGFP-WIPI2 IDR (364-425aa)Stable expression and inducible recruitment of protein in mammalian cell culture
223759pCAG_mCherry-ATG13 HORMA (1-191aa)Expression of recombinant protein for purification
223760pGEX4T1_GST-EGFP-ATG13_IDR (191-517aa)Expression of recombinant protein for purification
223761pGEX4T1_GST-TEV-EGFP-ATG13_IDR_191-517aa_(M196A_S197A_R199A)Expression of recombinant protein for purification
223762pETDuet1_6xHis-TEV-mCherry-ATG13 IDR (191-517aa)Expression of recombinant protein for purification
223763pETDuet1_6xHis-TEV-mCherry-WIPI3Expression of recombinant protein for purification
223764pFastBac-Dual_BNIP3(1-158aa)-THROMBIN-GSTExpression of recombinant protein for purification
223765pFastBac-Dual_BNIP3(1-158aa)-EGFP-TEV-GSTExpression of recombinant protein for purification
223766pFastBac-Dual_BNIP3(1-158aa)-EGFP-TEV-GST (W18A/L21A; deltaLIR)Expression of recombinant protein for purification
223767pHAGE-FKBP-mEGFP-WIPI1 Stable expression and inducible recruitment of protein in mammalian cell culture
223768pHAGE-FKBP-mEGFP-WIPI3 Stable expression and inducible recruitment of protein in mammalian cell culture
223769pHAGE-FKBP-mEGFP-WIPI4Stable expression and inducible recruitment of protein in mammalian cell culture
223770pHAGE-FKBP-mEGFP-WIPI2d (R108E/R125E)Stable expression and inducible recruitment of protein in mammalian cell culture
223771pGenlenti_ATG13-WTStable expression of protein in mammalian cell culture, rescue experiments
223772pGenlenti_ATG13-delta191-230Stable expression of protein in mammalian cell culture, rescue experiments
223773pGenlenti_ATG13-delta191-205Stable expression of protein in mammalian cell culture, rescue experiments
223774pGenlenti_ATG13-delta-206-230Stable expression of protein in mammalian cell culture, rescue experiments
223775pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (I224A/L227A; deltaLIR3)Expression of recombinant protein for purification
223776pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (I307A/V310A; deltaLIR4)Expression of recombinant protein for purification
223777pFastBac-Dual_BNIP3(1-158aa)-THROMBIN-GST (E44A L47A D49A A50K Q51A; deltaWIPI2)Expression of recombinant protein for purification
223778pFastBac-Dual_BNIP3(1-158aa)-THROMBIN-GST (W18A/L21A; deltaLIR)Expression of recombinant protein for purification
223779pGEX4T1_GST-EGFP-ATG13_IDR_191-517aa_(M196-H214 11xA)Expression of recombinant protein for purification
223780pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (I224A/L227A/W276A/I279A; deltaLIR1+3)Expression of recombinant protein for purification
223781pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (W276A/I279A/I307A/V310A; deltaLIR1+4)Expression of recombinant protein for purification
223782pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (Y213A/I216A/W276A/I279A; deltaLIR1+2)Expression of recombinant protein for purification
223783pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (Y213A/I216A; deltaLIR2)Expression of recombinant protein for purification
223784pETDuet1_BCL2L13(1-465aa)-EGFP-TEV-GST (I224A/L227A/W276A/I279A/I307A/V310A; deltaLIR1+3+4)Expression of recombinant protein for purification
223785pGEX4T1_GST-EGFP-ATG13_IDR_delta191-230 (231-517aa)Expression of recombinant protein for purification
223786pGEX4T1_GST-EGFP-ATG13_IDR_delta191-204 (205-517aa)Expression of recombinant protein for purification
223787pGEX4T1_GST-EGFP-ATG13_IDR_delta206-230 (191-205_231-517aa)Expression of recombinant protein for purification
223788pGenLenti V5-NIX (W36A/L39A; deltaLIR)Stable expression of protein in mammalian cell culture
223789pGenLenti V5-NIX (E72A/L75A/D77A/E81A; deltaWIPI2)Stable expression of protein in mammalian cell culture
223790pETDuet1_6xHis-TEV-mCherry-WIPI2d IDR (364-425aa)Expression of recombinant protein for purification
223791pGEX-4T1_GST-EGFP-ATG13_IDR (191-205aa)Expression of recombinant protein for purification
223792pGEX-4T1_GST-EGFP-ATG13_IDR (191-230aa)Expression of recombinant protein for purification
223793pGEX-4T1_GST-EGFP-ATG13_IDR (206-230aa)Expression of recombinant protein for purification
223794pHAGE_FKBP-mEGFP-WIPI2d_P2A-FRB-Fis1 (no mt-mKeima)Stable expression of protein in mammalian cell culture
223795pHAGE-FRB-Fis1 (no mt-mKeima)Stable expression of protein in mammalian cell culture
223796pCAG_ATG101Expression of recombinant protein for purification (co-expression with ATG13)
223797pCAG_GST-TEV-EGFP-ATG13Expression of recombinant protein for purification
223798pCAG_GST-TEV-WIPI1Expression of recombinant protein for purification
223799pCAG_WIPI2d-TEV-GSTExpression of recombinant protein for purification
223800pCAG_GST-TEV-WIPI3Expression of recombinant protein for purification
223801pCAG_GST-TEV-WIPI4Expression of recombinant protein for purification

Antibodies from Article