• Inicio
  • Resultados de la búsqueda
  • Large-scale label-free comparative proteomics analysis of polo-like kinase 1 inhibition via the small-molecule inhibitor BI 6727 (Volasertib) in BRAF(V600E) mutant melanoma cells.

Large-scale label-free comparative proteomics analysis of polo-like kinase 1 inhibition via the small-molecule inhibitor BI 6727 (Volasertib) in BRAF(V600E) mutant melanoma cells.

Journal of proteome research (2014-06-03)
Brian D Cholewa, Molly C Pellitteri-Hahn, Cameron O Scarlett, Nihal Ahmad
RESUMEN

Polo-like kinase 1 (Plk1) is a serine/threonine kinase that plays a key role during the cell cycle by regulating mitotic entry, progression, and exit. Plk1 is overexpressed in a variety of human cancers and is essential to sustained oncogenic proliferation, thus making Plk1 an attractive therapeutic target. However, the clinical efficacy of Plk1 inhibition has not emulated the preclinical success, stressing an urgent need for a better understanding of Plk1 signaling. This study addresses that need by utilizing a quantitative proteomics strategy to compare the proteome of BRAF(V600E) mutant melanoma cells following treatment with the Plk1-specific inhibitor BI 6727. Employing label-free nano-LC-MS/MS technology on a Q-exactive followed by SIEVE processing, we identified more than 20 proteins of interest, many of which have not been previously associated with Plk1 signaling. Here we report the down-regulation of multiple metabolic proteins with an associated decrease in cellular metabolism, as assessed by lactate and NAD levels. Furthermore, we have also identified the down-regulation of multiple proteasomal subunits, resulting in a significant decrease in 20S proteasome activity. Additionally, we have identified a novel association between Plk1 and p53 through heterogeneous ribonucleoprotein C1/C2 (hnRNPC), thus providing valuable insight into Plk1's role in cancer cell survival.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥98.5% (GC)
Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥99.0% (T)
Sigma-Aldrich
Monoclonal Anti-Human IgG (Fc specific) antibody produced in mouse, clone GG-7, ascites fluid
Sigma-Aldrich
Monoclonal Anti-Human IgG (Fc specific) antibody produced in mouse, clone HP-6017, ascites fluid
Sigma-Aldrich
Anti-Human IgG (Fc specific)−Biotin antibody, Mouse monoclonal, clone HP-6017, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Goat/Sheep IgG−Peroxidase antibody, Mouse monoclonal antibody produced in mouse, clone GT-34, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Rabbit IgG (γ-chain specific)−Alkaline Phosphatase antibody produced in mouse, clone RG-96, purified immunoglobulin, buffered aqueous glycerol solution
Sigma-Aldrich
Anti-Rabbit IgG (γ-chain specific)–Peroxidase antibody, Mouse monoclonal, clone RG-96, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Rabbit IgG (γ-chain specific) antibody produced in mouse, clone RG-96, ascites fluid
Sigma-Aldrich
Anti-Human IgG (Fc specific)−FITC antibody, Mouse monoclonal antibody produced in mouse, clone HP-6017, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Goat/Sheep IgG antibody produced in mouse, clone GT-34, ascites fluid
Sigma-Aldrich
Anti-Rabbit IgG, Native−Peroxidase antibody, Mouse monoclonal antibody produced in mouse, clone RabT-50, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Human IgG (Fab specific) antibody produced in mouse, clone GG-6, ascites fluid
Sigma-Aldrich
Anti-Rabbit IgG, Native antibody, Mouse monoclonal, clone RabT-50, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Rabbit IgG (γ-chain specific)−Biotin antibody, Mouse monoclonal, clone RG-96, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Human IgG2 (Fc specific) antibody produced in mouse, clone HP-6002, ascites fluid
Sigma-Aldrich
Anti-Goat/Sheep IgG−Biotin antibody, Mouse monoclonal, clone GT-34, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Goat/Sheep IgG−Alkaline Phosphatase antibody, Mouse monoclonal, clone GT-34, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Goat/Sheep IgG−FITC antibody, Mouse monoclonal antibody produced in mouse, clone GT-34, purified from hybridoma cell culture
Sigma-Aldrich
Monoclonal Anti-Human IgG4 (pFc′ specific) antibody produced in mouse, clone HP-6023, ascites fluid
Sigma-Aldrich
Monoclonal Anti-Human IgG (γ-chain specific) antibody produced in mouse, clone GG-5, ascites fluid
Sigma-Aldrich
Monoclonal Anti-Rat IgG2a antibody produced in mouse, clone R2A-2, ascites fluid