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1 St. Vincent's Institute, Fitzroy, Australia
2 Protein Chemistry and Metabolism, St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia
* To whom correspondence should be addressed. E-mail: mwatt{at}svi.edu.au.
We have investigated the gene and protein expression of ATGL and TG lipase activity from subcutaneous and visceral adipose tissue of lean and obese subjects. Visceral and subcutaneous adipose tissue was obtained from 16 age-matched lean and obese subjects during abdominal surgery. Tissues were analysed for mRNA expression of lipolytic enzymes by real time quantitative-PCR. ATGL protein content was assessed by western blot and TG lipase activity by radiometric assessment. Subcutaneous and visceral adipose tissue of obese subjects had elevated mRNA expression of PNPLA2 (ATGL), and other lipases including PNPLA3, PNPLA4, CES1 and LYPLAL1 (P<0.05). Surprisingly, ATGL protein expression and TG lipase activity were reduced in subcutaneous adipose tissue of obese subjects. Immunoprecipitation of ATGL reduced total TG lipase activity in adipose lysates by 70% in obese and 83% in lean subjects. No significant difference in the ATGL activator CGI-58 mRNA levels (ABHD5) were associated with obesity. These data demonstrate that ATGL is important for efficient TG lipase activity in humans. They also demonstrate reduced ATGL protein expression and TG lipase activity despite increased mRNA expression of ATGL and other novel lipolytic enzymes in obesity. The lack of correlation between ATGL protein content and in vitro TG lipase activity indicates that small decrements in ATGL protein expression are not responsible for the reduction in TG lipase activity observed here in obesity and that posttranslational modifications may be important.
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