Apolipoprotein B-containing lipoproteins (B-lps) are essential for the transport of hydrophobic dietary and endogenous lipids through the circulation in vertebrates. Zebrafish embryos produce large numbers of B-lps in the yolk syncytial layer (YSL) to move lipids from yolk to growing tissues. Disruptions in B-lp production perturb yolk morphology, readily allowing for visual identification of mutants with altered B-lp metabolism. Here we report the discovery of a missense mutation in microsomal triglyceride transfer protein (Mtp), a protein that is essential for B-lp production. This mutation of a conserved glycine residue to valine (zebrafish G863V, human G865V) reduces B-lp production and results in yolk opacity due to aberrant accumulation of cytoplasmic lipid droplets in the YSL. However, this phenotype is milder than that of the previously reported L475P stalactite ( stl ) mutation. MTP transfers lipids, including triglycerides and phospholipids, to apolipoprotein B in the ER for B-lp assembly. In vitro lipid transfer assays reveal that while both MTP mutations eliminate triglyceride transfer activity, the G863V mutant protein unexpectedly retains ~80% of phospholipid transfer activity. This residual phospholipid transfer activity of the G863V mttp mutant protein is sufficient to support the secretion of small B-lps, which prevents intestinal fat malabsorption and growth defects observed in the mttp stl/stl mutant zebrafish. Modeling based on the recent crystal structure of the heterodimeric human MTP complex suggests the G865V mutation may block triglyceride entry into the lipid-binding cavity. Together, these data argue that selective inhibition of MTP triglyceride transfer activity may be a feasible therapeutic approach to treat dyslipidemia and provide structural insight for drug design. These data also highlight the power of yolk transport studies to identify proteins critical for B-lp biology. Author summary Cardiovascular disease affects about a third of the world’s population and is mediated by the buildup of lipids (primarily derived from plasma lipoproteins) in the wall of blood vessels. An essential protein required for the synthesis of the lipoproteins, from insects to humans, is microsomal triglyceride transfer protein (MTP). This protein transfers lipids, including triglycerides and phospholipids, to apolipoprotein B (APOB) for the assembly of ApoB-containing lipoproteins. Here we report a mutation in MTP that blocks the transfer of triglycerides but not phospholipids. Modeling based on the recent crystal structure of the MTP complex suggests the G865V mutation may block access to the region of the protein that binds triglyceride. In zebrafish, the residual phospholipid transfer activity of the mutant protein is sufficient to support secretion of small ApoB-containing lipoproteins and prevent a number of serious health conditions observed in humans harboring null MTP mutations (e.g., intestinal fat malabsorption, growth retardation). These results suggest that selective inhibitors of MTP that can mimic this mutation may be a feasible therapeutic approach to treat dyslipidemias in humans.
【초록키워드】 Mutation, drug design, Lipoproteins, Human, cardiovascular disease, metabolism, cardiovascular, Protein, humans, blood vessels, modeling, Triglyceride, Apolipoprotein B, phenotype, Missense mutation, mutants, morphology, mutant, circulation, Insects, lipids, crystal structure, inhibitor, synthesis, disease, zebrafish, Critical, dyslipidemia, assembly, Phospholipids, Therapeutic approach, power, Lipid, triglycerides, Disruption, growth retardation, glycine, growth defects, valine, growth retardation), selective inhibition, Support, Transport, ApoB, tissues, growth, These data, complex, opacity, residue, secretion, transfer, treat, hydrophobic, selective, wall, ApoB-containing lipoproteins, Apolipoprotein B-containing lipoproteins, B-lps, cytoplasmic lipid droplets, dyslipidemias, fat malabsorption, G863V mttp mutant protein, G863V mutant protein, intestinal fat malabsorption, microsomal triglyceride transfer protein, MTP complex, MTP triglyceride transfer activity, MTTP, plasma lipoproteins, vertebrates, yolk, yolk opacity, yolk syncytial layer, insect, health condition, block, Affect, Prevent, highlight, bind, Embryo, identify, conserved, reported, required, reduce, feasible, ApoB-containing lipoprotein, Apolipoprotein B-containing lipoprotein, B-lp, cytoplasmic lipid droplet, endogenous lipid, G863V, heterodimeric, human MTP, L475P, Mtp, YSL, 【제목키워드】 Protein, Point mutation, lipid transfer activity,