title: Purification, cellular levels, and functional domains of lipase maturation factor 1.
abstract: Over a third of the US adult population has hypertriglyceridemia, resulting in an increased risk of atherosclerosis, pancreatitis, and metabolic syndrome. Lipoprotein lipase (LPL), a dimeric enzyme, is the main lipase responsible for TG clearance from the blood after food intake. LPL requires an endoplasmic reticulum (ER)-resident, transmembrane protein known as lipase maturation factor 1 (LMF1) for secretion and enzymatic activity. LMF1 is believed to act as a client specific chaperone for dimeric lipases, but the precise mechanism by which LMF1 functions is not understood. Here, we examine which domains of LMF1 contribute to dimeric lipase maturation by assessing the function of truncation variants. N-terminal truncations of LMF1 show that all the domains are necessary for LPL maturation. Fluorescence microscopy and protease protection assays confirmed that these variants were properly oriented in the ER. We measured cellular levels of LMF1 and found that it is expressed at low levels and each molecule of LMF1 promotes the maturation of 50 or more molecules of LPL. Thus we provide evidence for the critical role of the N-terminus of LMF1 for the maturation of LPL and relevant ratio of chaperone to substrate.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States.
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Babilonia-Rosa
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, United States. Electronic address: neher@email.unc.edu.
authors.forename: Saskia B
authors.initials: SB
authors.lastname: Neher
mesh_terms: D000595:Amino Acid Sequence; D004789:Enzyme Activation; D006801:Humans; D008071:Lipoprotein Lipase / Q000737:chemistry* / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000737:chemistry* / Q000378:metabolism*
chemical_list: C546366:LMF1 protein, human; D008565:Membrane Proteins; D008071:Lipoprotein Lipase
keywords: Cellular protein levels; Chaperone; Endoplasmic reticulum; Lipase maturation factor 1; Lipid metabolism; Lipoprotein lipase