Characterization of a Splice Variant of Perilipin 5

Abstract

Lipid storage provides energy stores for the body in times of increased oxidative requirements, such as fasting or exercise, but when the balance of lipid storage and mobilization is disrupted human health suffers. Implications for increased lipid storage include risk factors for developing type II diabetes mellitus and non‐alcoholic fatty liver disease. The perilipin family of proteins associate with the surface of lipid droplets and are essential to regulation of stored triacylglycerols. Perilipin 5 functions to maintain a balance between the storage of triacylglycerols in lipid droplets or their mobilization when energetically required. Several members of the perilipin gene family exhibit differential splicing, most notably perilipin 1 but reports exist of alternative messages for shortened forms of perilipins 2 and 3. We have identified a shortened form of perilipin 5 we have termed perilipin 5b. Western blots indicate that the protein, of approximately 35 kDa, is present in C2C12 cultured myoblasts and in heart and liver tissue from fed and fasted mice. Analysis of murine genomic DNA sequences reveals stop codons found in the introns between exons 6 and 7 and 7 and 8, either of which could give rise to a protein of approximately 35 kDa, indicating the likelihood of the observed protein being a splice variant, rather than a post‐translationally modified protein. Using RT‐PCR we have identified multiple amplicons suggesting that perilipin‐5b contains a read through of intron 6 of the murine perilipin 5 gene. Analysis of the amino acid sequence of perilipin 5b indicates that this truncation would lack the reported C‐terminal domain essential for interactions with ATGL and CGI‐58 but would retain the PKA site found at Ser155. The function of perilipin 5b is yet unknown, but the fact that it is found primarily in heart tissue seems to suggest that it plays a role in the oxidation of the lipid droplet. These data are consistent with what is currently known about Perilipin‐5 and the other PAT family proteins. Determination of perilipin 5b function will give a more accurate picture of the overall mechanism of lipid storage and metabolism.