Recombinant Human MCL1, His-tagged

• Cat.No.: MCL1-779H
• Product Overview: Recombinant Human MCL1 protein, fused to His-tag, was expressed in E.coli and purified by Ni-sepharose.

Specification

• Species: Human
• Source: E.coli
• Tag: His
• Protein Length: 1-350aa
• Storage: The protein is stored in PBS buffer at -20℃. Avoid repeated freezing and thawing cycles.
• Storage Buffer: 1M PBS (58mM Na2HPO4,17mM NaH2PO4, 68mM NaCl, pH8. ) added with 300mM Imidazole and 0.7% Sarcosyl, 15%glycerol.

Gene Information

• Gene Name: MCL1 myeloid cell leukemia sequence 1 (BCL2-related) [ Homo sapiens ]
• Official Symbol: MCL1
• Synonyms: MCL1; myeloid cell leukemia sequence 1 (BCL2-related); induced myeloid leukemia cell differentiation protein Mcl-1; BCL2L3; Mcl 1; bcl-2-like protein 3; myeloid cell leukemia ES; bcl-2-related protein EAT/mcl1; TM; EAT; MCL1L; MCL1S; Mcl-1; MCL1-ES; bcl2-L-3; mcl1/EAT; MGC1839; MGC104264
• Gene ID: 4170
• mRNA Refseq: NM_001197320
• Protein Refseq: NP_001184249
• MIM: 159552
• UniProt ID: Q07820
• Chromosome Location: 1q21
• Pathway: Apoptosis, organism-specific biosystem; Direct p53 effectors, organism-specific biosystem; E2F transcription factor network, organism-specific biosystem; HIF-1-alpha transcription factor network, organism-specific biosystem; IL-7 Signaling Pathway, organism-specific biosystem; IL6-mediated signaling events, organism-specific biosystem
• Function: BH3 domain binding; protein binding; protein channel activity; protein heterodimerization activity

Citation

Mcl-1 interacts with Akt to promote lung cancer progression

Journal: Cancer research    PubMed ID: 31662324    Data: 2020/6/15

Authors: Guo Chen, Dongkyoo Park, Xingming Deng

Article Snippet:BrdU cell proliferation kit and purified recombinant His tag-Akt protein (#14–276) were purchased from EMD Millipore (Billerica, MA).BrdU cell proliferation kit and purified recombinant His tag-Akt protein (#14–276) were purchased from EMD Millipore (Billerica, MA).. Purified recombinant His-tag-Mcl-1 protein (MCL1–781H) was obtained from Creative BioMart (Shirley, NY).. Mcl-1 rabbit polyclonal Ab (sc-819), anti-β-actin (sc-47778), anti-Bax (sc-493), anti-HA (sc-7392) and anti-GFP (sc-9996) antibodies as well as Akt1/2 siRNA (sc-43609) were purchased from Santa Cruz Biotechnology (Dallas, Texas).Mcl-1 rabbit polyclonal Ab (sc-819), anti-β-actin (sc-47778), anti-Bax (sc-493), anti-HA (sc-7392) and anti-GFP (sc-9996) antibodies as well as Akt1/2 siRNA (sc-43609) were purchased from Santa Cruz Biotechnology (Dallas, Texas).

Knockout of Mcl-1 suppresses cancer cell growth via downregulation of Akt activity. A-E, Mcl-1 was knocked out from H1299 cells using CRISPR/Cas9, followed by analysis of Mcl-1 expression by Western blot (A), growth curve (B), colony formation (C), human phospho-kinase array (1: Akt S473; 2: Akt T308) (D), and quantification of human phospho-kinase by image J (E). Data represent the mean ± SD. *P < 0.05, ***P < 0.001, by 2-tailed t test. (F) Mcl-1 was knocked out from H1299 cells, or knocked down from H460 cells, followed by Western blotting analysis of Mcl-1, pAkt, pmTOR and p-p70S6K.

The binding of Mcl-1 with Akt is essential for Mcl-1 to disrupt PH/KD interactions, upregulate Akt activity and promote tumor growth. A and B, GST-tagged kinase domain of Akt and GFP-tagged PH domain of Akt were co-transfected with Flag-tagged WT Mcl-1 or ΔPEST Mcl-1 mutant into H1299 cells, followed by GST pull down (A), or co-IP using GFP antibody (B), and Western blot using GST or GFP antibody. C, Interaction between the kinase domain and PH domain in the absence or presence of WT Mcl-1 or ΔPEST was measured in a mammalian two-hybrid system. D and E, H1299 Mcl-1?/? cells were transfected with empty vector (EV), Flag-tagged WT Mcl-1 or ΔPEST Mcl-1 mutant, followed by analysis of pAkt, pmTOR and p-p70 S6K by Western blot (D) and colony formation assay (E). Data represent the mean ± SD. ***P < 0.001, by 2-tailed t test. F, The same number (3×106) of H1299 Mcl-1?/? cells expressing EV, Flag-tagged WT Mcl-1 or ΔPEST mutant were injected into subcutaneous tissue in the flank region of nude mice to generate lung cancer xenografts. Tumor volume was measured once every 3 days. After 33 days, the mice were sacrificed and the tumors were removed, photographed, and analyzed. Data represent the mean ± SD, n=6 mice per group. ***P < 0.001, by 2-tailed t test.

Knockout of Mcl-1 inhibits tumor growth in vivo. A and B, The same number (3×106) of H1299 parental or Mcl-1?/? cells were injected into subcutaneous tissue in the flank region of nude mice to generate lung cancer xenografts (n=5 mice each group). Tumor volume was measured once every 3 days. After 33 days, the mice were sacrificed and the tumors were removed, photographed, and analyzed. Data represent the mean ± SD, n=5 per group. ***P < 0.001, by 2-tailed t test. C-H, IHC staining of Mcl-1, pAkt (S473), pAkt (T308), Ki67 and active caspase 3 was performed in tumor tissues at the end of experiments. Data represent the mean ± SD, n=5 per group. **P < 0.01, ***P < 0.001, by 2-tailed t test.

First evidence that oligopyridines, α-helix foldamers, inhibit Mcl-1 and sensitize ovarian carcinoma cells to Bcl-xL-targeting strategies.

Journal: Journal of medicinal chemistry    PubMed ID: 25585174    Data: 2015/5/7

Authors: Céline Gloaguen, Anne Sophie Voisin-Chiret, Laurent Poulain

Article Snippet:Capture of His-Bcl-xL (same condition as His-Mcl-1) was performed on channel Fc1 that was used as a reference surface for nonspecific binding measurements.Capture of His-Bcl-xL (same condition as His-Mcl-1) was performed on channel Fc1 that was used as a reference surface for nonspecific binding measurements.. Recombinant human Mcl-1 protein, fused to His-tag, and recombinant human Bcl-xL, His-tagged (DNA sequence encoding the human BCL2L1 isoform 1 (Met 1?Arg 212) fused with a polyhistidine tag at the C-terminus) were purchased from Creative BioMart (45?16 Ramsey Road, Shirley, NY 11967, USA).. A lowmass weight-single-cycle kinetics (LMW-SCK) analysis to determine association, dissociation, and affinity constants (ka, kd, and Kd respectively) was carried out by injecting different protein concentrations.A lowmass weight-single-cycle kinetics (LMW-SCK) analysis to determine association, dissociation, and affinity constants (ka, kd, and Kd respectively) was carried out by injecting different protein concentrations.