Regulators of TNFα Mediated Insulin Resistance Elucidated by Quantitative Proteomics

Abstract

Obesity is a growing epidemic worldwide and is a major risk factor for several chronic diseases, including diabetes, kidney disease, heart disease, and cancer. Obesity often leads to type 2 diabetes mellitus (T2DM, via the increased production of proinflammatory cytokines such as tumor necrosis factor-α (TNFα). Our study combines different proteomic techniques to investigate the changes in the global proteome, secretome and phosphoproteome of adipocytes under chronic inflammation condition, as well as fundamental cross-talks between different cellular pathways regulated by chronic TNFα exposure. Our results show that many key regulator proteins of the canonical and non-canonical NF-κB pathways, such as Nfkb2, and its downstream effectors, including Csf-1 and Lgals3bp, directly involved in leukocyte migration and invasion, were significantly upregulated at the intra and extracellular levels, culminating in the progression of inflammation. Our data provides evidence of several key proteins that play a role in the development of insulin resistance.

Caveolin‐2 in association with nuclear lamina controls adipocyte hypertrophy

“…We then explored Cav‐2 stability in three experimental obese adipocyte models. Briefly, normal mature adipocytes differentiated up to 8 d in adipogenic induction medium (AIM) (N) were further incubated in the post‐differentiation medium (PDM) alone for 8 and 16 d (P‐8, P‐16: Obese adipocytes 33,34 ), or with insulin (PI‐8, PI‐16: Obese adipocytes with excessive lipogenesis 35,36 ) or tumor necrosis factor α (TNFα) (PT‐8, PT‐16: Obese adipocytes with insulin resistance 36,37 ). Obese adipocytes (Ob) were then selected by measuring an expansion of lipid droplet area as compared to the normal adipocytes (Figure 3D).…”

“…adipocytes with insulin resistance 36,37 ). Obese adipocytes were then selected by measuring an expansion of lipid droplet area as compared to the normal adipocytes (Figure 3D).…”

Expression profiles of an inactive aspartic protease (Bla g 2 allergen) in different tissues and developmental stages of the German cockroach (Blattella germanica)

“…Peptides were resuspended in 10μl of 3% ACN/0.1% Formic Acid (FA)/96.9% MilliQ, and 5 μl was used for LC‐MS/MS analysis Peptides were analyzed in the Dionex UltiMate 3000 RSLC nano System coupled to the Q‐Exactive High‐Field (HF) Hybrid Quadrupole Orbitrap MS (Thermo Fisher Scientific) as described elsewhere (Connelly et al, 2018; Mohallem & Aryal, 2020; Zembroski et al, 2021) using 130‐min LC method. The MS data were acquired with a Top 20 data‐dependent MS/MS scan method with a maximum injection time of 100 ms, a resolution of 120,000 at 200 m/z .…”

Legionella pneumophila temporally regulates the activity of ADP/ATP translocases by reversible ADP‐ribosylation

“…Protein bands were digested in-gel with Lys-C as previously described 67 . Digested peptides were analyzed by LC-ESI-MS/MS using the Dionex UltiMate 3000 RSLC nano System coupled to the Q Exactive™ HF Hybrid Quadrupole-Orbitrap Mass Spectrometer (Thermo Fisher Scientific) as described previously 68,69 . Reverse-phase peptide separation was performed using a trap column (300 μm ID × 5 mm) packed with 5 μm 100 Å PepMap C18 medium, and then separated on a reverse-phase column (50 cm long × 75 µm ID) packed with 2 µm 100 Å PepMap C18 silica (Thermo Fisher Scientific).…”