Alkenylhydrolase: A Microsomal Enzyme Activity in Rat Brain

Rat brain microsomes have the capacity to liberate radioactive free aldehydes from 1‐[1‐ 14 C]alk‐1′‐enyl‐ sn ‐glycero‐3‐phosphoethanolamine (lysoplasmalogen). Glycerophosphoethanolamine was found using 1‐alk‐1′‐enyl‐ sn ‐glycero‐3‐phospho‐[ 3 H]ethanolamine. The ratio of both products released by lysoplasmalogenase action was 1:1. Another enzymic activity could be demonstrated, which hydrolyzes lysoplasmalogen at the hydrophilic part of the molecule, a lysophospholipid phospho diesterase. Thus, 1‐[1‐ 14 C]alk‐1′‐enylglycerol was detected as well as [ 3 H]ethanolamine, again in a molar ratio, from the respective labeled substrates. This enzyme possesses nearly the same affinity toward the substrate as lysoplasmalogenase. Whereas the lysophospholipid phos phodiesterase is totally inhibited in the presence of NaF or EDTA, lysoplasmalogenase activity is not affected by these reagents. 1‐[1‐‘ 14 C]Alk‐l’‐enylglycerol acts also as substrate for lysoplasmalogenase, which liberates radioactive aldehydes at the same rate as from lysoplasmal ogen. Because the apparent K m and V max values are nearly identical for both substrates, the enzyme activities are inhibited in the same way, and the pH optimum is about 7.2 in both cases, it is concluded that both substrates were attacked by the same enzyme. The enzyme does not differentiate between a substrate substituted at the sn ‐3 position of glycerol and one that is not. It requires only a free OH group at the sn ‐2 position. Phosphoethanolamine phosphatase activity was also determined under our experimental conditions.