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ampka 1 2  (Cell Signaling Technology Inc)
94
Cell Signaling Technology Inc ampka 1 2
Figure 3. SUMOylation Promotes LKB1-AMPK Interaction (A) In vivo assay in HeLa cells of binding between AMPK and LKB1 wild-type or K178R mutant after 6 hr of metabolic stress is shown. (B) Stably LKB1-expressing A549 cells were transiently transfected with either empty vector (pcDNA3) or FLAG SENP1 followed by treatment for 20 hr with vehicle (CAMKK inhibitor) alone or with 2 mM phenformin. LKB1 was immunoprecipitated from lysates and their interaction with AMPK was assessed. See also Figure S3A. (C) Co-immunoprecipitation assay of endogenous AMPK and HA-SUMO1 in A549 cells is shown. (D) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the absence or presence of PIASy SIM peptides is shown. (E) The AMPK domain is shown (green; PDB: 4CFH). Highlighted are the following: inhibitory staurosporin (white carbons), V93 (gray), I94 (orange), phosphor- ylated T172 (yellow carbons), and the activation loop (orange). (Top right) Zoom onto I94 shows that I94 is mostly solvent exposed, suggesting that its mutation to alanine is unlikely to compromise the 3D fold or stability of the AMPK domain. (Bottom right) The non-covalent recognition of SUMO1 (blue) by RanBP2 (green, only the interacting part is shown) through b strand interactions (PDB: 3UIP) was used as a structural basis for the search for potential SIMs in AMPK. The di- hydrophobic motif figuring a valine and isoleucine are shown. The AMPK N-terminal b sheet offers a surface similar to a potential SIM (top right). (F) A schematic shows a conserved region in the N-terminal region of AMPK that conforms to the specifications of a previously proposed SIM (Minty et al., 2000). (G) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the presence of wild-type or mutant AMPK SIM peptides is shown. (H) Peptide pull-down assay of biotinylated wild-type or mutant AMPK SIM peptides conjugated to avidin beads in HEK293 cell lysates is shown. (I and J) In vivo assay in <t>AMPKa/</t> MEF cells of binding between AMPK or AMPK SIM mutants and (I) SUMO1 and (J) LKB1 after 6 hr of metabolic stress. Whole- cell lysates were subjected to western blot using the indicated antibodies.
Ampka 1 2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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protective effects against siv attacks  (ATCC)
97
ATCC protective effects against siv attacks
Figure 3. SUMOylation Promotes LKB1-AMPK Interaction (A) In vivo assay in HeLa cells of binding between AMPK and LKB1 wild-type or K178R mutant after 6 hr of metabolic stress is shown. (B) Stably LKB1-expressing A549 cells were transiently transfected with either empty vector (pcDNA3) or FLAG SENP1 followed by treatment for 20 hr with vehicle (CAMKK inhibitor) alone or with 2 mM phenformin. LKB1 was immunoprecipitated from lysates and their interaction with AMPK was assessed. See also Figure S3A. (C) Co-immunoprecipitation assay of endogenous AMPK and HA-SUMO1 in A549 cells is shown. (D) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the absence or presence of PIASy SIM peptides is shown. (E) The AMPK domain is shown (green; PDB: 4CFH). Highlighted are the following: inhibitory staurosporin (white carbons), V93 (gray), I94 (orange), phosphor- ylated T172 (yellow carbons), and the activation loop (orange). (Top right) Zoom onto I94 shows that I94 is mostly solvent exposed, suggesting that its mutation to alanine is unlikely to compromise the 3D fold or stability of the AMPK domain. (Bottom right) The non-covalent recognition of SUMO1 (blue) by RanBP2 (green, only the interacting part is shown) through b strand interactions (PDB: 3UIP) was used as a structural basis for the search for potential SIMs in AMPK. The di- hydrophobic motif figuring a valine and isoleucine are shown. The AMPK N-terminal b sheet offers a surface similar to a potential SIM (top right). (F) A schematic shows a conserved region in the N-terminal region of AMPK that conforms to the specifications of a previously proposed SIM (Minty et al., 2000). (G) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the presence of wild-type or mutant AMPK SIM peptides is shown. (H) Peptide pull-down assay of biotinylated wild-type or mutant AMPK SIM peptides conjugated to avidin beads in HEK293 cell lysates is shown. (I and J) In vivo assay in <t>AMPKa/</t> MEF cells of binding between AMPK or AMPK SIM mutants and (I) SUMO1 and (J) LKB1 after 6 hr of metabolic stress. Whole- cell lysates were subjected to western blot using the indicated antibodies.
Protective Effects Against Siv Attacks, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/in+vitro+analysis+(siva)+tool/Lactiplantibacillus+plantarum/pm39101975-169-164-174
Average 97 stars, based on 1 article reviews
protective effects against siv attacks - by Bioz Stars, 2026-09
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Image Search Results


Figure 3. SUMOylation Promotes LKB1-AMPK Interaction (A) In vivo assay in HeLa cells of binding between AMPK and LKB1 wild-type or K178R mutant after 6 hr of metabolic stress is shown. (B) Stably LKB1-expressing A549 cells were transiently transfected with either empty vector (pcDNA3) or FLAG SENP1 followed by treatment for 20 hr with vehicle (CAMKK inhibitor) alone or with 2 mM phenformin. LKB1 was immunoprecipitated from lysates and their interaction with AMPK was assessed. See also Figure S3A. (C) Co-immunoprecipitation assay of endogenous AMPK and HA-SUMO1 in A549 cells is shown. (D) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the absence or presence of PIASy SIM peptides is shown. (E) The AMPK domain is shown (green; PDB: 4CFH). Highlighted are the following: inhibitory staurosporin (white carbons), V93 (gray), I94 (orange), phosphor- ylated T172 (yellow carbons), and the activation loop (orange). (Top right) Zoom onto I94 shows that I94 is mostly solvent exposed, suggesting that its mutation to alanine is unlikely to compromise the 3D fold or stability of the AMPK domain. (Bottom right) The non-covalent recognition of SUMO1 (blue) by RanBP2 (green, only the interacting part is shown) through b strand interactions (PDB: 3UIP) was used as a structural basis for the search for potential SIMs in AMPK. The di- hydrophobic motif figuring a valine and isoleucine are shown. The AMPK N-terminal b sheet offers a surface similar to a potential SIM (top right). (F) A schematic shows a conserved region in the N-terminal region of AMPK that conforms to the specifications of a previously proposed SIM (Minty et al., 2000). (G) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the presence of wild-type or mutant AMPK SIM peptides is shown. (H) Peptide pull-down assay of biotinylated wild-type or mutant AMPK SIM peptides conjugated to avidin beads in HEK293 cell lysates is shown. (I and J) In vivo assay in AMPKa/ MEF cells of binding between AMPK or AMPK SIM mutants and (I) SUMO1 and (J) LKB1 after 6 hr of metabolic stress. Whole- cell lysates were subjected to western blot using the indicated antibodies.

Journal: Cell reports

Article Title: A Critical SUMO1 Modification of LKB1 Regulates AMPK Activity during Energy Stress.

doi: 10.1016/j.celrep.2015.07.002

Figure Lengend Snippet: Figure 3. SUMOylation Promotes LKB1-AMPK Interaction (A) In vivo assay in HeLa cells of binding between AMPK and LKB1 wild-type or K178R mutant after 6 hr of metabolic stress is shown. (B) Stably LKB1-expressing A549 cells were transiently transfected with either empty vector (pcDNA3) or FLAG SENP1 followed by treatment for 20 hr with vehicle (CAMKK inhibitor) alone or with 2 mM phenformin. LKB1 was immunoprecipitated from lysates and their interaction with AMPK was assessed. See also Figure S3A. (C) Co-immunoprecipitation assay of endogenous AMPK and HA-SUMO1 in A549 cells is shown. (D) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the absence or presence of PIASy SIM peptides is shown. (E) The AMPK domain is shown (green; PDB: 4CFH). Highlighted are the following: inhibitory staurosporin (white carbons), V93 (gray), I94 (orange), phosphor- ylated T172 (yellow carbons), and the activation loop (orange). (Top right) Zoom onto I94 shows that I94 is mostly solvent exposed, suggesting that its mutation to alanine is unlikely to compromise the 3D fold or stability of the AMPK domain. (Bottom right) The non-covalent recognition of SUMO1 (blue) by RanBP2 (green, only the interacting part is shown) through b strand interactions (PDB: 3UIP) was used as a structural basis for the search for potential SIMs in AMPK. The di- hydrophobic motif figuring a valine and isoleucine are shown. The AMPK N-terminal b sheet offers a surface similar to a potential SIM (top right). (F) A schematic shows a conserved region in the N-terminal region of AMPK that conforms to the specifications of a previously proposed SIM (Minty et al., 2000). (G) In vitro SUMO1 pull-down assay of GST-AMPKA1 recombinant protein in the presence of wild-type or mutant AMPK SIM peptides is shown. (H) Peptide pull-down assay of biotinylated wild-type or mutant AMPK SIM peptides conjugated to avidin beads in HEK293 cell lysates is shown. (I and J) In vivo assay in AMPKa/ MEF cells of binding between AMPK or AMPK SIM mutants and (I) SUMO1 and (J) LKB1 after 6 hr of metabolic stress. Whole- cell lysates were subjected to western blot using the indicated antibodies.

Article Snippet: Antibodies and siRNAs Antibodies used for immunoblotting, including LKB1 (D60C5; 3047), phosphoAMPK Thr172 (2531), total AMPKa 1/2 (2532), phospho-Raptor Ser792 (2083), total Raptor (2280), phospho-ULK1 (S555; 5869), total ULK1 (8054), and SUMO1 (4930), were obtained from Cell Signaling Technology.

Techniques: In Vivo, Binding Assay, Mutagenesis, Stable Transfection, Expressing, Transfection, Plasmid Preparation, Immunoprecipitation, Co-Immunoprecipitation Assay, In Vitro, Pull Down Assay, Recombinant, Activation Assay, Solvent, Avidin-Biotin Assay, Western Blot