Selective inhibitors of the glycosylphosphatidylinositol biosynthetic pathway of Trypanosoma brucei

TY - JOUR

T1 - Selective inhibitors of the glycosylphosphatidylinositol biosynthetic pathway of Trypanosoma brucei

AU - Smith, Terry K.

AU - Sharma, Deepak K.

AU - Crossman, Arthur

AU - Brimacombe, John S.

AU - Ferguson, Michael A. J.

PY - 1999/11/1

Y1 - 1999/11/1

N2 - Synthetic analogues of D-GlcN alpha 1-6D-myo-inositol-1-HPO4-3(sn-1,2-diacylglycerol) (GlcN-PI), with the 2-position of the inositol residue substituted with an O-octyl ether [D-GlcN alpha 1-6D-(2-O-octyl)myo-inositol-1-HPO4-3-sn-1,2-dipalmitoylglycerol; GlcN-(2-O-octyl) PI] or O-hexadecyl ether [D-GlcN alpha 1-6D-(2-O-hexadecyl)myo-inositol-1-HPO4-3-sn-1,2-dipalmitoylglycerol; GlcN-(2-O-hexadecyl)PI], were tested as substrates or inhibitors of glycosylphosphatidylinositol (GPI) biosynthetic pathways using cell-free systems of the protozoan parasite Trypanosoma brucei (the causative agent of human African sleeping sickness) and human HeLa cells. Neither these compounds nor their N-acetyl derivatives are substrates or inhibitors of GPI biosynthetic enzymes in the HeLa cell-free system but are potent inhibitors of GPI biosynthesis in the T.brucei cell-free system. GlcN-(2-O-hexadecyl)PI was shown to inhibit the first alpha-mannosyltransferase of the trypanosomal GPI pathway. The N-acetylated derivative GlcNAc-(2-O-octyl)PI is a substrate for the trypanosomal GlcNAc-PI de-N-acetylase and this compound, like GlcN-(2-O-octyl)PI, is processed predominantly to Man(2)GlcN-(2-O-octyl)PI by the T.brucei cell-free system. Both GlcN-(2-O-octyl)PI and GlcNAc(2-O-octyl)PI also inhibit inositol acylation of Man(1-3)GlcN-PI and, consequently, the addition of the ethanolamine phosphate bridge in the T.brucei cell-free system. The data establish these substrate analogues as the first generation of in vitro parasite GPI pathway-specific inhibitors.

AB - Synthetic analogues of D-GlcN alpha 1-6D-myo-inositol-1-HPO4-3(sn-1,2-diacylglycerol) (GlcN-PI), with the 2-position of the inositol residue substituted with an O-octyl ether [D-GlcN alpha 1-6D-(2-O-octyl)myo-inositol-1-HPO4-3-sn-1,2-dipalmitoylglycerol; GlcN-(2-O-octyl) PI] or O-hexadecyl ether [D-GlcN alpha 1-6D-(2-O-hexadecyl)myo-inositol-1-HPO4-3-sn-1,2-dipalmitoylglycerol; GlcN-(2-O-hexadecyl)PI], were tested as substrates or inhibitors of glycosylphosphatidylinositol (GPI) biosynthetic pathways using cell-free systems of the protozoan parasite Trypanosoma brucei (the causative agent of human African sleeping sickness) and human HeLa cells. Neither these compounds nor their N-acetyl derivatives are substrates or inhibitors of GPI biosynthetic enzymes in the HeLa cell-free system but are potent inhibitors of GPI biosynthesis in the T.brucei cell-free system. GlcN-(2-O-hexadecyl)PI was shown to inhibit the first alpha-mannosyltransferase of the trypanosomal GPI pathway. The N-acetylated derivative GlcNAc-(2-O-octyl)PI is a substrate for the trypanosomal GlcNAc-PI de-N-acetylase and this compound, like GlcN-(2-O-octyl)PI, is processed predominantly to Man(2)GlcN-(2-O-octyl)PI by the T.brucei cell-free system. Both GlcN-(2-O-octyl)PI and GlcNAc(2-O-octyl)PI also inhibit inositol acylation of Man(1-3)GlcN-PI and, consequently, the addition of the ethanolamine phosphate bridge in the T.brucei cell-free system. The data establish these substrate analogues as the first generation of in vitro parasite GPI pathway-specific inhibitors.

KW - Glycosylphosphatidylinositol (GPI)

KW - HeLa

KW - Inositol acyltransferase

KW - Mannosyltransferase

KW - Trypanosoma

U2 - 10.1093/emboj/18.21.5922

DO - 10.1093/emboj/18.21.5922

M3 - Article

C2 - 10545104

SN - 0261-4189

VL - 18

SP - 5922

EP - 5930

JO - EMBO Journal

JF - EMBO Journal

IS - 21

ER -