| First Author | Yin F | Year | 2009 |
| Journal | J Lipid Res | Volume | 50 |
| Issue | 11 | Pages | 2265-9 |
| PubMed ID | 19471043 | Mgi Jnum | J:154958 |
| Mgi Id | MGI:4411976 | Doi | 10.1194/jlr.M900196-JLR200 |
| Citation | Yin F, et al. (2009) A quantitative assay measuring the function of lipase maturation factor 1. J Lipid Res 50(11):2265-9 |
| abstractText | Newly synthesized lipoprotein lipase (LPL) and related members of the lipase gene family require an endoplasmic reticulum maturation factor for attainment of enzyme activity. This factor has been identified as lipase maturation factor 1 (Lmf1), and mutations affecting its function and/or expression result in combined lipase deficiency (cld) and hypertriglyceridemia. To assess the functional impact of Lmf1 sequence variations, both naturally occurring and induced, we report the development of a cell-based assay using LPL activity as a quantitative reporter of Lmf1 function. The assay uses a cell line homozygous for the cld mutation, which renders endogenous Lmf1 nonfunctional. LPL transfected into the mutant cld cell line fails to attain activity; however, cotransfection of LPL with wild-type Lmf1 restores its ability to support normal lipase maturation. In this report, we describe optimized conditions that ensure the detection of a complete range of Lmf1 function (full, partial, or complete loss of function) using LPL activity as the quantitative reporter. To illustrate the dynamic range of the assay, we tested several novel mutations in mouse Lmf1. Our results demonstrate the ability of the assay to detect and analyze Lmf1 mutations having a wide range of effects on Lmf1 function and protein expression. |
