Identification of covalently attached fatty acids in the hydrophobic membrane-binding domain of human erythrocyte acetylcholinesterase.

Human erythrocyte acetylcholinesterase is an amphipathic enzyme whose hydrophobic membrane-binding domain can be selectively labeled with a lipophilic photoreagent and removed by digestion with papain. In this paper we demonstrate that methanolysis releases covalently bound fatty acids from the hydrophobic domain and thus confirm that this domain is a covalently linked glycolipid at the enzyme subunit C-terminus. About one mole of saturated and one mole of unsaturated fatty acids were released per mole of domain. Since the predominant unsaturated fatty acids (22:4 and 22:5) are minor components of the esterified fatty acid pool in human erythrocyte membranes, assembly of the glycolipid must involve a selected unsaturated fatty acid pool.

[1]  K. Waku,et al.  Distinctive selectivity for docosatetraenoic acid incorporation by Ehrlich ascites tumor cells. , 1982, Biochimica et biophysica acta.

[2]  I. Silman,et al.  Physicochemical behaviour and structural characteristics of membrane-bound acetylcholinesterase from Torpedo electric organ. Effect of phosphatidylinositol-specific phospholipase C. , 1985, The Biochemical journal.

[3]  D. M. Scoggin,et al.  Structure of human erythrocyte acetylcholinesterase. Characterization of intersubunit disulfide bonding and detergent interaction. , 1984, The Journal of biological chemistry.

[4]  N. Tattrie Positional distribution of saturated and unsaturated fatty acids on egg lecithin , 1959 .

[5]  R. G. Ackman,et al.  Application of specific response factors in the gas chromatographic analysis of methyl esters of fatty acids with flame ionization detectors , 1964 .

[6]  C. Klee,et al.  Identification of the NH2‐terminal blocking group of calcineurin B as myristic acid , 1982, FEBS letters.

[7]  I. Silman,et al.  A hydrophobic dimer of acetylcholinesterase from Torpedo californica electric organ is solubilized by phosphatidylinositol-specific phospholipase C , 1983, Neuroscience Letters.

[8]  T. Rosenberry,et al.  A small hydrophobic domain that localizes human erythrocyte acetylcholinesterase in liposomal membranes is cleaved by papain digestion. , 1985, Biochemistry.

[9]  K. Haldar,et al.  Trypanosoma brucei variant surface glycoprotein has a sn-1,2-dimyristyl glycerol membrane anchor at its COOH terminus. , 1985, The Journal of biological chemistry.

[10]  M. G. Low,et al.  Non‐lytic release of acetylcholinesterase from erythrocytes by A phosphatidylinositol‐specific phospholipase C , 1977, FEBS letters.

[11]  G. Cross,et al.  Glycopeptides from variant surface glycoproteins of Trypanosoma Brucei. C-terminal location of antigenically cross-reacting carbohydrate moieties. , 1981, Molecular and biochemical parasitology.

[12]  I Silman,et al.  Identification of covalently bound inositol in the hydrophobic membrane-anchoring domain of Torpedo acetylcholinesterase. , 1985, Biochemical and biophysical research communications.

[13]  G. Cross Structure of the variant glycoproteins and surface coat of Trypanosoma brucei. , 1984, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[14]  T. Baltz,et al.  Presence of glycerol and fatty acids in the C-terminal end of a variant surface glycoprotein from Trypanosoma equiperdum. , 1983, Biochemical and biophysical research communications.

[15]  W. Ruitenbeek The fatty acid composition of various lipid fractions isolated from erythrocytes and blood plasma of patients with Duchenne and congenital myotonic muscular dystrophy. , 1978, Clinica chimica acta; international journal of clinical chemistry.

[16]  D. Campbell,et al.  Rat brain Thy-1 glycoprotein. The amino acid sequence, disulphide bonds and an unusual hydrophobic region. , 1981, The Biochemical journal.

[17]  T. Rosenberry,et al.  Human erythrocyte acetylcholinesterase is an amphipathic protein whose short membrane-binding domain is removed by papain digestion. , 1984, The Journal of biological chemistry.

[18]  G. Kolata Novel protein/membrane attachment. , 1985, Science.

[19]  A. Holder Carbohydrate is linked through ethanolamine to the C-terminal amino acid of Trypanosoma brucei variant surface glycoprotein. , 1983, The Biochemical journal.