BACKGROUND: Apolipoprotein A5 (APOA5) over-expression enhances lipolysis of triglyceride (TG) through stimulation of lipoprotein lipase (LPL) activity; however, an APOA5 G185C variant was found associated with hypertriglyceridemia. The aim of this study was, therefore, to explore the importance of APOA5 185GG in the activation of LPL. METHODS: A fragment containing mature human APOA5 cDNA was obtained by RT-PCR and subcloned into pET-15b vector. Site-directed mutagenesis was performed to generate 19 variants. Recombinant human APOA5 wild type and variants were produced in Escherichia coli, and then activation of LPL was measured. RESULTS: Activity of APOA5 variants on LPL-mediated 1,2-dimyristoyl-sn-glycero-3-phosphocholine hydrolysis was reduced by 17 to 74% in comparison to wild type APOA5 (P<0.0001). All variants also showed reduced activation (P<0.0001) of LPL-mediated hydrolysis of very low-density lipoprotein (VLDL); activation abilities of APOA5 variants ranged from 31 to 81% of wild-type APOA5. CONCLUSIONS: APOA5 residue 185G is very important in LPL-mediated VLDL hydrolysis, and any mutation at this residue will decrease LPL activation and concomitant TG modulation.
BACKGROUND:Apolipoprotein A5 (APOA5) over-expression enhances lipolysis of triglyceride (TG) through stimulation of lipoprotein lipase (LPL) activity; however, an APOA5G185C variant was found associated with hypertriglyceridemia. The aim of this study was, therefore, to explore the importance of APOA5 185GG in the activation of LPL. METHODS: A fragment containing mature humanAPOA5 cDNA was obtained by RT-PCR and subcloned into pET-15b vector. Site-directed mutagenesis was performed to generate 19 variants. Recombinant humanAPOA5 wild type and variants were produced in Escherichia coli, and then activation of LPL was measured. RESULTS: Activity of APOA5 variants on LPL-mediated 1,2-dimyristoyl-sn-glycero-3-phosphocholine hydrolysis was reduced by 17 to 74% in comparison to wild type APOA5 (P<0.0001). All variants also showed reduced activation (P<0.0001) of LPL-mediated hydrolysis of very low-density lipoprotein (VLDL); activation abilities of APOA5 variants ranged from 31 to 81% of wild-type APOA5. CONCLUSIONS:APOA5 residue 185G is very important in LPL-mediated VLDL hydrolysis, and any mutation at this residue will decrease LPL activation and concomitant TG modulation.
Authors: Ying Wu; Amanda F Marvelle; Jin Li; Damien C Croteau-Chonka; Alan B Feranil; Christopher W Kuzawa; Yun Li; Linda S Adair; Karen L Mohlke Journal: J Lipid Res Date: 2013-09-10 Impact factor: 5.922
Authors: Vineeta Sharma; Andrzej Witkowski; H Ewa Witkowska; Andrew Dykstra; Jens B Simonsen; Lisa Nelbach; Jennifer A Beckstead; Clive R Pullinger; John P Kane; Mary J Malloy; Gordon Watson; Trudy M Forte; Robert O Ryan Journal: Arterioscler Thromb Vasc Biol Date: 2014-08-14 Impact factor: 8.311
Authors: Elena Mendoza-Barberá; Josep Julve; Stefan K Nilsson; Aivar Lookene; Jesús M Martín-Campos; Rosa Roig; Alfonso M Lechuga-Sancho; John H Sloan; Pablo Fuentes-Prior; Francisco Blanco-Vaca Journal: J Lipid Res Date: 2013-01-10 Impact factor: 5.922
Authors: João Paulo G Camporez; Shoichi Kanda; Max C Petersen; François R Jornayvaz; Varman T Samuel; Sanjay Bhanot; Kitt Falk Petersen; Michael J Jurczak; Gerald I Shulman Journal: J Lipid Res Date: 2014-12-29 Impact factor: 5.922
Authors: Ying Wu; Lindsay L Waite; Anne U Jackson; Wayne H-H Sheu; Steven Buyske; Devin Absher; Donna K Arnett; Eric Boerwinkle; Lori L Bonnycastle; Cara L Carty; Iona Cheng; Barbara Cochran; Damien C Croteau-Chonka; Logan Dumitrescu; Charles B Eaton; Nora Franceschini; Xiuqing Guo; Brian E Henderson; Lucia A Hindorff; Eric Kim; Leena Kinnunen; Pirjo Komulainen; Wen-Jane Lee; Loic Le Marchand; Yi Lin; Jaana Lindström; Oddgeir Lingaas-Holmen; Sabrina L Mitchell; Narisu Narisu; Jennifer G Robinson; Fred Schumacher; Alena Stančáková; Jouko Sundvall; Yun-Ju Sung; Amy J Swift; Wen-Chang Wang; Lynne Wilkens; Tom Wilsgaard; Alicia M Young; Linda S Adair; Christie M Ballantyne; Petra Bůžková; Aravinda Chakravarti; Francis S Collins; David Duggan; Alan B Feranil; Low-Tone Ho; Yi-Jen Hung; Steven C Hunt; Kristian Hveem; Jyh-Ming J Juang; Antero Y Kesäniemi; Johanna Kuusisto; Markku Laakso; Timo A Lakka; I-Te Lee; Mark F Leppert; Tara C Matise; Leena Moilanen; Inger Njølstad; Ulrike Peters; Thomas Quertermous; Rainer Rauramaa; Jerome I Rotter; Jouko Saramies; Jaakko Tuomilehto; Matti Uusitupa; Tzung-Dau Wang; Michael Boehnke; Christopher A Haiman; Yii-Der I Chen; Charles Kooperberg; Themistocles L Assimes; Dana C Crawford; Chao A Hsiung; Kari E North; Karen L Mohlke Journal: PLoS Genet Date: 2013-03-21 Impact factor: 5.917