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. 2021 Aug 10;10:e71188. doi: 10.7554/eLife.71188

Figure 5. Native mass spectrometry indicates the presence of coenzyme A in the mutant TACAN sample.

(A, B) Mass species detected in purified wild-type (A) and His196Ala, His197Ala mutant (B) TACAN protein without treatment (“untreated”), or incubated with CoASH (MW = 767.5 Da), S-ethyl-CoA (MW = 795.6 Da), or acetyl-CoA (MW = 809.6 Da).

Figure 5.

Figure 5—figure supplement 1. TACAN is co-purified with coenzyme A molecules.

Figure 5—figure supplement 1.

(A) Binding details of CoASH in TACANH196A H197A. Surface charge is represented from blue (positive charges) to red (negative charges). Neighboring residues are shown as sticks, and the hydrogen bonding is indicated by a black dash. A CoASH molecule is shown as sticks and colored according to atom type. (B) Conformation comparison of eicosanol-CoA in ELOVL7 (green) and CoASH in TACAN (blue) based on the alignment in Figure 4A. (C) CoASH releasing activity of ELOVL7 and TACAN. Proteoliposomes of TACAN and ELOVL7 reconstituted in soy L-α-phosphatidylcholine (soy-PC) at 1:50 protein-to-lipid ratio (w/w) with 10 μg protein and 500 μg soy-PC were used. Empty proteoliposomes made of soy-PC were used as control. ELOVL7 showed significant activity, while neither the wild-type nor His196Ala His197Ala mutant of TACAN showed any activity.