A study was conducted to determine whether retinyl phosphate would act as substrate for the enzymatic synthesis of mannosyl retinyl phosphate. Retinyl phosphate, prepared chemically, supported the growth of vitamin A deficient rats at the same rate as retinol. It also stimulated the uptake of [14C]mannose from GDP [14C]mannose into total chloroform methanol extractable lipid. This reaction occurred in the presence of ATP, Mn2+, detergent (Zonyl A), and a membrane rich enzyme preparation from the livers of vitamin A deficient rats, provided that a lipid extract of the membrane preparation of α L lecithin was also added. Total chloroform methanol extractable, labeled mannolipid was separated into 2 principal labeled mannolipids by thin layer or column chromatography or by differential solvent extraction. The properties of these mannolipids identified them as glycophospholipids: one was identical with authentic synthetic dolichyl mannosyl phosphate, and the other was concluded to be mannosyl retinyl phosphate because of its incorporation of radioactivity from [3H]retinyl phosphate, its rapid hydrolysis by dilute acid, and the formation of a substance that cochromatographed with retinol upon its acid hydrolysis. The presence of ATP or GTP was essential for the stimulation of mannolipid synthesis, probably because of their protective action on the substrates against phosphatases present in the crude enzyme fraction. A pH of 6.0-6.2 favored the formation of dolichyl mannosyl phosphate; a higher pH (6.7-7.0) that of mannosyl retinyl phosphate.
|Original language||English (US)|
|Number of pages||9|
|Journal||Journal of Lipid Research|
|State||Published - Dec 1 1975|
ASJC Scopus subject areas
- Cell Biology