Functional Assessment of Perilipin 3 Lipid Binding Domains with Stearic Acid
Yeomans, Michael M.
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In 2011–2014, the prevalence of obesity was just over 36% in adults and 17% in youth. The rising numbers of overweight and obese individuals has lead to an increase in the number of associated chronic disease like nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). These diseases are characterized by hepatic lipid accumulation and increased inflammation, conditions that are closely associated with the primary hepatic lipid droplet (LD) protein perilipin 2 (Plin2). Prior research with Plin2 has found it plays a role in hepatic lipid regulation; Plin2 stands to serve as a valuable protein with regard to future therapeutic and treatment options for chronic diseases associated with obesity. To better distinguish Plin2’s unique role and characteristics within the Perilipin Family of proteins, research on its closest analog perilipin 3 (Plin3) is warranted. A better understanding of the perilipin family of proteins could result in more effective and efficient treatment options for diseases related to obesity. The following study was conducted to assess (1) a new method of expression and purification for use with Plin2 capitalizing on an N-terminal His6-SUMO protein fusion; (2) the lipid binding domain of Plin3 and several of its substitution mutant proteins via fluorescence binding to serve as a basis for comparison for future research conducted with Plin2 and similar mutants. Major findings of this study indicate (A) success with Plin3 protein fusion expression and purification that warrants the method’s use with Plin2; (B) significantly greater affinity for stearic ligands amongst the Plin3 substitution mutant proteins as compared to the WT and to one another. Thus, the current work provides valuable information for future research with Plin2 and the rest of the perilipin family.