Characterization of human SCD2, an oligomeric desaturase with improved stability and enzyme activity by cross-linking in intact cells.
SCD (stearoyl-CoA desaturase) catalyses the conversion of saturated fatty acids into mono-unsaturated fatty acids, a critical step involved in lipid metabolism and various other biological functions. In the present study, we report the identification and characterization of a human gene that encodes a novel SCD enzyme (hSCD2). The hSCD2 gene codes for a 37.5-kDa protein that shares 61% and 57% sequence identity with the human SCD1 and mouse SCD2 enzymes respectively. The recombinant hSCD2 enzyme expressed in mammalian and Sf9 insect cells efficiently catalysed desaturation of both stearoyl- and palmitoyl-CoAs to the corresponding mono-unsaturated fatty acids. In comparison with the hSCD1 gene that is predominantly expressed in liver, hSCD2 is most abundantly expressed in pancreas and brain. Additionally, hSCD2 transcripts from adult and foetal tissues exhibit different sizes because of alternative splicing in the non-coding region, suggesting that hSCD2 expression is developmentally regulated. The recombinant human SCD2 and SCD1 transiently expressed in COS-7 cells exhibited as oligomeric proteins that consist of homodimers and oligomers when resolved by SDS/PAGE. The complex formation was independent of SCD protein expression levels, as supported by a relatively constant ratio of the level of dimers and oligomers to that of the monomers from COS-7 cells transiently transfected with different amounts of SCD expression vectors. Furthermore, treatment of intact COS-7 cells with a cross-linking reagent resulted in dose-dependent increases in the levels of SCD protein and activity, suggesting that oligomerization may play an important role in regulating the stability of SCD enzymes.
Zhang
Shaobo
S
Endocrine Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
Yang
Yanzhu
Y
Shi
Yuguang
Y
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology; D002460:Cell Line; D044943:Fatty Acid Desaturases / Q000737:chemistry* / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology; D010920:Placenta / Q000201:enzymology; D011485:Protein Binding; D021122:Protein Subunits; D017386:Sequence Homology, Amino Acid; D013230:Stearoyl-CoA Desaturase / Q000737:chemistry* / Q000378:metabolism*; D014018:Tissue Distribution
D021122:Protein Subunits; D044943:Fatty Acid Desaturases; D013230:Stearoyl-CoA Desaturase