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Functional Characterization Reveals Human ELOVL3 Role in the Biosynthesis of iso ‐20:0
Author(s) -
Wang Zhen,
Park Hui Gyu,
Lawrence Peter,
Kothapalli Kumar S. D.,
Brenna J. Thomas
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.684.13
Subject(s) - trypsinization , transfection , fatty acid , substrate (aquarium) , chemistry , biochemistry , incubation , biosynthesis , biology , gene , trypsin , enzyme , ecology
Branched chain fatty acids (BCFA) are major, bioactive components of the human diet, present from later fetal life throughout normal aging. Our objective was to investigate the catalytic activity of saturated fatty acid elongases ( ELOVL1, ELOVL3, ELOVL6 and ELOVL7 ) by checking for enhanced conversion of BCFA substrate to product. Methods The open reading frame of ELOVL1, 3, 6 and 7 were cloned into a pcDNA3.1 expression vector. ELOVL1, ELOVL3, ELOVL6, ELOVL7 and empty vector control were transiently transfected into MCF7 breast cancer cells using Polyplus jetPRIME transfection reagent. Twenty four hours (h) after transfection cells were supplemented with 80 μM of albumin‐bound iso ‐18:0 fatty acid substrate and were allowed to incubate for 24 h. After incubation, the cells were washed twice with 1× PBS, removed by trypsinization and fatty acids analyzed. Results ELOVL3 transfected cells showed enhanced conversion of iso ‐18:0 to iso ‐20:0 compared to control and other elongases. The relative conversions of iso ‐18:0 → iso ‐20:0 from two biological replicates for ELOVL3 and control was 3.96 ± 0.01 and 2.02 ± 0.02, respectively. ELOVL7 showed moderate activity (3.14 ± 0.16), whereas, ELOVL1 and ELOVL6 showed no activity towards iso ‐18:0. Conclusion This is the first report of human elongase(s) showing activity specifically towards BCFA substrate. Support or Funding Information NIH R01 AT007003