Glucolipid Synthesis in *Acanthamoeba castellanii**

HAROLD B. SKRDLANT,

HAROLD B. SKRDLANT

Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas 78284

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ROBERT A. WEISMAN,

ROBERT A. WEISMAN

Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas 78284

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HAROLD B. SKRDLANT,

HAROLD B. SKRDLANT

Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas 78284

Search for more papers by this author

ROBERT A. WEISMAN,

ROBERT A. WEISMAN

Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas 78284

Search for more papers by this author

First published: November 1976

https://doi.org/10.1111/j.1550-7408.1976.tb03852.x

^†

This investigation was supported by Research Grant AI 10241 from NIAID, U.S. Public Health Service and Research Grant AQ 419, from the Robert A. Welch Foundation.

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Abstract

SYNOPSIS. Cell-free preparations of Acanthamoeba castellanii trophozoites transfer glucose from UDP-[U-¹⁴C]glucose to a chloroform-soluble form. This radioactive material has been isolated by thin-layer chromatography; it contains an alkali-labile and an alkali-stable (unsaponifiable) component. Treatment of the enzymic product with 0.1 N KOH for 15 min at 0 C or 20 C releases radioactivity into the aqueous phase as glucose. During this treatment, 30–60% of the original glycolipid remains chloroform-soluble. It is considered to be an alkali-stable glycolipid because no further loss of radioactivity occurs during an additional 45-min of treatment with 0.1 N KOH. During incubation with 0.1 N HCI at 100 C glucose is released quantitatively from both the untreated glycolipid and the alkali-stable glycolipid with a half-time of 6 min. Glycolipid formation is inhibited by UDP and is reversible; extracts catalyze the formation of UDP-glucose from the alkali-stable glucolipid and UDP.

The chemical and physical properties of the alkali-stable glycolipid are consistent with a glucosyl phosphoryl polyprenol structure. Extracts prepared from cysts catalyze the formation of glycolipids aiso, but the glucosyltransferase activity/cell decreases during the course of encystment. Radioactivity is incorporated into the fraction insoluble in chloroform-methanol-water (1:1:1:) during these incubations when UDP-[U-¹⁴C]glucose or [¹⁴C]glycolipid is the substrate.

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Volume23, Issue4

November 1976

Pages 613-618

Glucolipid Synthesis in *Acanthamoeba castellanii**

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Glucolipid Synthesis in Acanthamoeba castellanii*

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Glucolipid Synthesis in *Acanthamoeba castellanii**