In many species, the lactating mammary gland is one of the most lipogenic organs of the body. The majority of the lipid produced during lactation is secreted into milk by a novel process of membrane envelopment of cytoplasmic lipid droplets (CLDs). Adipophilin (ADRP/ADPH/PLIN2), a member of the perilipin (PAT) family of lipid droplet proteins, is hypothesized to play a pivotal role in both formation and secretion of milk lipids. Production of milk lipids is the only known example of CLD secretion, and the only process in which PAT family members undergo secretion. This review discusses emerging data about the structural and functional properties of adipophilin that determine its physiological actions and mediate its effects on milk lipid formation and secretion.

Cytoplasmic lipid droplets (CLD) in mammary epithelial cells undergo secretion by a unique membrane envelopment process to produce milk lipids. Adipophilin (ADPH/Plin2), a member of the perilipin/PAT family of lipid droplet-associated proteins, is hypothesized to mediate CLD secretion through interactions with apical plasma membrane elements. We found that the secretion of CLD coated by truncated ADPH lacking the C-terminal region encoding a putative four-helix bundle structure was impaired relative to that of CLD coated by full-length ADPH. We used homology modeling and analyses of the solution and membrane binding properties of purified recombinant ADPH C terminus to understand how this region possibly mediates CLD secretion. Homology modeling supports the concept that the ADPH C terminus forms a four-helix bundle motif and suggests that this structure can form stable membrane bilayer interactions. Circular dichroism and protease mapping studies confirmed that the ADPH C terminus is an independently folding ␣-helical structure that is relatively resistant to urea denaturation. Liposome binding studies showed that the purified C terminus binds to phospholipid membranes through electrostatic dependent interactions, and cell culture studies documented that it localizes to the plasma membrane. Collectively, these data provide direct evidence that the ADPH C terminus forms a stable membrane binding helical structure that is important for CLD secretion. We speculate that interactions between the four-helix bundle of ADPH and membrane phospholipids may be an initial step in milk lipid secretion.Milk lipids are an essential source of neonatal calories and provide nutrients in the form of fatty acids and bioactive lipids that are required for growth and development (1). Milk lipids are delivered to the newborn as milk fat globules (MFG) 4 derived from triglyceride-rich cytoplasmic lipid droplets (CLD) (2). Adipophilin (ADPH/ADRP/Adfp/Plin2), a member of the perilipin/PAT (perilipin/adipophilin/TIP47) family of proteins (3), is an abundant MFG protein (4 -6) and a prominent CLDassociated protein of mammary epithelial cells (7,8). PAT proteins have been shown to promote the formation and accumulation of CLD in a variety of tissues (9, 10) and are also implicated in the trafficking and secretion of CLD (2, 9). The lactating mammary gland is an ideal system to study the recurrent cycles of CLD formation, accumulation, and secretion, and previous work from our laboratory has implicated ADPH in each of these processes (5,8,11).CLD are secreted during lactation by an apocrine process where the apical plasma membrane engulfs associated CLD, which are then released by a budding process to form MFG (2). Two key regulators of CLD secretion are the transmembrane protein butyrophilin (BTN) and the cytoplasmic homodimer, xanthine oxidoreductase (XOR). Mice deficient in either of these proteins display impaired CLD envelopment and secretion (12, 13). In addition to being an abundant protein on mammary epithelial CLD and secreted MFG ...