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. 1978 Mar;61(3):357–360. doi: 10.1104/pp.61.3.357

Subcellular Distribution of Steryl Ester Biosynthesis in Spinach Leaves

Raymond E Garcia 1,1, J Brian Mudd 1
PMCID: PMC1091867  PMID: 16660292

Abstract

Higher steryl ester biosynthetic activities were obtained with Triton X-100-phosphatidylcholine-cholesterol mixed micelles than with Tween 80-phosphatidylcholine-cholesterol mixed micelles when incubated with spinach leaf (Spinacia oleracea L.) acetone powder preparations. The best incorporation of [4-14C]cholesterol into [4-14C]cholesteryl ester was obtained with a Triton X-100-phosphatidylcholine-cholesterol (10:1:1, w/w) mixed micelle system. This mixed micelle system, however, required 1,2-dipalmitin and fatty acid-free bovine albumin for optimal activity. The reaction exhibited a diglyceride specificity since the dipalmitin requirement could be replaced with neither 1-monopalmitin nor tripalmitin. Significant amounts of steryl ester biosynthetic activity were detected in the chloroplast (1,000g pellet), mitochondrial (3,000g pellet), and microsomal (20,000g and 88,000g pellet) fractions. Little activity was detected in the water-soluble (88,000g supernatant) fraction. The highest specific activity occurred in the 88,000g pellet. The 88,000g supernatant contained a heatstable, water-soluble substance that was required for optimal steryl ester biosynthesis in all of the pellet fractions. This factor was not lost during extensive dialysis but was destroyed by ashing, indicating that it was large and organic. Silver nitrate thin layer chromatography indicated that 60% of the biosynthesized steryl esters contained saturated fatty acids in the absence of 1,2-dipalmitin and that 83% contained saturated fatty acids in the presence of 1,2-dipalmitin.

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Selected References

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