Mature adipocyte proteome reveals differentially altered protein abundances between lean, overweight and morbidly obese human subjects

Benabdelkamel, Hicham; Almidani, Ghaith M.; Alsadhan, A; Bassas, Abdulelah F.; Duncan, Mark W.; Alfadda, Assim A.

doi:10.1016/j.mce.2014.11.021

Cited by 37 publications

(67 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Actins B and G, that were significantly decreased in the VAT of MUHO group compared to the MHO group, are also known to participate in the expansion capability of the ECM (Alfadda et al, 2013; van Vliet-Ostaptchouk et al, 2014). The function and role of ANXA5, ACTG, ACTB, and LEG1 have been shown in our previous proteomic studies (Alfadda et al, 2013; Benabdelkamel et al, 2015). …”

Section: Discussionsupporting

confidence: 58%

“…The suspension was shaken for 1 h at room temperature and then sonicated (Microsonicator, Qsonica Sonicators, USA; 30% pulse, two intervals of 1 min each, separated by a 1 min gap). 50mM dithiothreitol (DTT) was then added and the protein extracts were centrifuged (20,000 × g, 1 h, 4°C) according to the procedure developed by our group (Alfadda et al, 2013; Benabdelkamel et al, 2015). The pellet was removed and the solubilized protein in the supernatant was precipitated using the 2D Clean-up Kit according to the manufacturer’s protocol (GE Healthcare, USA).…”

Section: Methodsmentioning

confidence: 99%

“…This was denatured in lysis buffer and mixed with a rehydration buffer. Gels were stained with Colloidal Coomassie Blue as described previously (Alfadda et al, 2013; Benabdelkamel et al, 2015). …”

Section: Methodsmentioning

confidence: 99%

“…Coomassie stained gel spots were excised manually, then washed and digested according to the previously described protocol (Benabdelkamel et al, 2015). In brief, destaining was carried out in steps, first by adding 50 mM NH 4 HCO 3 and 50% CH 3 CN, then by adding 100% CH 3 CN.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

A Proteomics Based Approach Reveals Differential Regulation of Visceral Adipose Tissue Proteins between Metabolically Healthy and Unhealthy Obese Patients

Alfadda

Masood

Al-Naami

et al. 2017

Molecules and Cells

View full text Add to dashboard Cite

Obesity and the metabolic disorders that constitute metabolic syndrome are a primary cause of morbidity and mortality in the world. Nonetheless, the changes in the proteins and the underlying molecular pathways involved in the relevant pathogenesis are poorly understood.In this study a proteomic analysis of the visceral adipose tissue isolated from metabolically healthy and unhealthy obese patients was used to identify presence of altered pathway(s) leading to metabolic dysfunction. Samples were obtained from 18 obese patients undergoing bariatric surgery and were subdivided into two groups based on the presence or absence of comorbidities as defined by the International Diabetes Federation. Two dimensional difference in-gel electrophoresis coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was carried out. A total of 28 proteins were identified with a statistically significant difference in abundance and a 1.5-fold change (ANOVA, p ≤ 0.05) between the groups. 11 proteins showed increased abundance while 17 proteins were decreased in the metabolically unhealthy obese compared to the healthy obese. The differentially expressed proteins belonged broadly to three functional categories: (i) protein and lipid metabolism (ii) cytoskeleton and (iii) regulation of other metabolic processes. Network analysis by Ingenuity pathway analysis identified the NFκB, IRK/MAPK and PKC as the nodes with the highest connections within the connectivity map. The top network pathway identified in our protein data set related to cellular movement, hematological system development and function, and immune cell trafficking. The VAT proteome between the two groups differed substantially between the groups which could potentially be the reason for metabolic dysfunction.

show abstract

Section: Discussionsupporting

confidence: 58%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A Proteomics Based Approach Reveals Differential Regulation of Visceral Adipose Tissue Proteins between Metabolically Healthy and Unhealthy Obese Patients

Alfadda

Masood

Al-Naami

et al. 2017

Molecules and Cells

View full text Add to dashboard Cite

show abstract

“…The floating layer of mature adipocytes was removed from the collagenase‐dispersed preparation and from the SVF pellet, washed three times in Dulbecco's modified eagle's medium, and stored at −80°C until analysis. The visceral adipose tissue samples were selected from a cohort of samples collected from patients undergoing bariatric surgery at the King Khalid University Hospital in Riyadh, Kingdom of Saudi Arabia . These samples were obtained from the greater omentum, immediately snap‐frozen in liquid nitrogen and stored at −80°C until analysis.…”

Section: Methodsmentioning

confidence: 99%

Comparison of protein precipitation methods from adipose tissue using difference gel electrophoresis

2018

Self Cite

View full text Add to dashboard Cite

Proteomic methods have great potential to aid our understanding of the functional and pathological roles of adipose tissue. A critical initial step in the proteomic studies is the efficient isolation of proteins before conducting detailed analysis. In this study, three different methods were used for precipitating proteins; we analyzed samples from visceral adipose tissue, subcutaneous adipose tissue, and stromal visceral fraction extracts after chloroform/methanol, acetone, and trichloroacetic acid precipitation. The proteins recovered after the precipitation steps were examined by 2D-DIGE. Statistical analyses were carried out using simple linear regression analyses and R2 values were calculated for the intra- and inter-method comparisons. We found that all three precipitation methods provided highly reproducible protein spots that were recovered when run in duplicate using the same method of precipitation, irrespective of whether it was solvent (R2 = 0.85-0.98) or acid-based (R2 = 0.80-0.96). A higher variation and poor correlation was noted for the recovered protein spots when comparisons were made between the methods (R2 = 0.40-0.88) and also when the same method was compared between different sample types. In this study, TCA-precipitated samples were enriched in lower molecular mass proteins compared to the samples extracted by solvent-based precipitation methods.

show abstract

Proteomic and Metabolite Profiling Reveals Profound Structural and Metabolic Reorganization of Adipocyte Mitochondria in Obesity

Schöttl

Pachl

Giesbertz

et al. 2020

Obesity

View full text Add to dashboard Cite

Objective Previous studies have revealed decreased mitochondrial respiration in adipocytes of obese mice. This study aimed to identify the molecular underpinnings of altered mitochondrial metabolism in adipocytes. Methods Untargeted proteomics of mitochondria isolated from adipocytes and metabolite profiling of adipose tissues were conducted in diet‐induced obese (DIO) and lean mice. Subcutaneous and intra‐abdominal adipose tissues were studied to depict depot‐specific alterations. Results In subcutaneous adipocytes of DIO mice, changes in proteins related to mitochondrial structure and function were observed. Mitochondrial proteins of the inner and outer membrane were strongly reduced, whereas proteins of key matrix metabolic pathways were increased in the obese versus lean state, as further substantiated by metabolite profiling. A pronounced decrease in the oxidative phosphorylation (OXPHOS) enzymatic equipment and cristae density of the inner membrane was identified. In intra‐abdominal adipocytes, similar systematic downregulation of the OXPHOS machinery in obesity occurred, but there was no regulation of outer membrane or matrix proteins. Conclusions Protein components of the OXPHOS machinery are systematically downregulated in adipose tissues of DIO mice compared with lean mice. Loss of the mitochondrial OXPHOS capacity in adipocytes may aggravate the development of metabolic disease.

show abstract

Mature adipocyte proteome reveals differentially altered protein abundances between lean, overweight and morbidly obese human subjects

Cited by 37 publications

References 61 publications

A Proteomics Based Approach Reveals Differential Regulation of Visceral Adipose Tissue Proteins between Metabolically Healthy and Unhealthy Obese Patients

A Proteomics Based Approach Reveals Differential Regulation of Visceral Adipose Tissue Proteins between Metabolically Healthy and Unhealthy Obese Patients

Comparison of protein precipitation methods from adipose tissue using difference gel electrophoresis

Proteomic and Metabolite Profiling Reveals Profound Structural and Metabolic Reorganization of Adipocyte Mitochondria in Obesity

Contact Info

Product

Resources

About