1 Ploug Group, BRIC Research Groups, BRIC, Københavns Universitet2 Chulalongkorn University, Bangkok, Thailand;3 University of California Los Angeles4 Cornell University, Ithaca, United States;5 Finsen Laboratory and BRIC, Copenhagen, Denmark;6 Umea University7 Rangsit University, Bangkok, Thailand.8 Ploug Group, BRIC Research Groups, BRIC, Københavns Universitet9 Umea University
GPIHBP1, a glycosylphosphatidylinositol-anchored glycoprotein of microvascular endothelial cells, binds lipoprotein lipase (LPL) within the interstitial spaces and transports it across endothelial cells to the capillary lumen. GPIHBP1's ability to bind LPL depends on its Ly6 domain, a three-fingered structure containing 10 cysteines and a conserved pattern of disulfide bond formation. Here, we report a patient with severe hypertriglyceridemia who was homozygous for a GPIHBP1 point mutation that converted a serine in GPIHBP1's Ly6 domain (Ser-107) to a cysteine. Two hypertriglyceridemic siblings were homozygous for the same mutation. All three homozygotes had very low levels of LPL in the pre-heparin plasma. We suspected that the extra cysteine in GPIHBP1-S107C might prevent the trafficking of the protein to the cell surface, but this was not the case. However, nearly all of the GPIHBP1-S107C on the cell surface was in the form of disulfide-linked dimers and multimers, while wild-type GPIHBP1 was predominantly monomeric. An insect cell GPIHBP1 expression system confirmed the propensity of GPIHBP1-S107C to form disulfide-linked dimers and to form multimers. Functional studies showed that only GPIHBP1 monomers bind LPL. In keeping with that finding, there was no binding of LPL to GPIHBP1-S107C in either cell-based or cell-free binding assays. We conclude that an extra cysteine in GPIHBP1's Ly6 motif results in multimerization of GPIHBP1, defective LPL binding, and severe hypertriglyceridemia.
Journal of Biological Chemistry, 2014, Vol 289, Issue 28, p. 19491-19499