Fig. 3.
A 4:1 mixture of oleic and erucic acids inhibits ELOVL1 by a mixed-type inhibition mechanism. The total membrane fraction (10 μg protein) from HeLa-ELOVL1 cells was incubated with the indicated concentration of C22:0-CoA and 0.075 μCi (27.3 μM) [2-14C]malonyl-CoA at 37°C for 30 min in the presence of 20 μM 4:1 mixture of oleic (C18:1) and erucic (C22:1) acids in ethanol or ethanol vehicle alone. After a sequential workup (see “Materials and Methods”), lipids were separated by normal-phase TLC and quantified by a bioimaging analyzer BAS-2500. (A) Values presented are the amount of FA generated per microgram of the total membrane protein. (B) The mean values from three independent experiments are expressed in a Lineweaver-Burk plot with a linear regression line. LO, 4:1 mixture of oleic and erucic acids.