Exploring the unique N-glycome of the opportunistic human pathogen Acanthamoeba.

TY - JOUR

T1 - Exploring the unique N-glycome of the opportunistic human pathogen Acanthamoeba.

AU - Schiller, Birgit

AU - Makrypidi, Georgia

AU - Razzazi-Fazeli, Ebrahim

AU - Paschinger, Katharina

AU - Walochnik, Julia

AU - Wilson, Iain B H

PY - 2012

Y1 - 2012

N2 - Glycans play key roles in host-pathogen interactions; thus, knowing the N-glycomic repertoire of a pathogen can be helpful in deciphering its methods of establishing and sustaining a disease. Therefore, we sought to elucidate the glycomic potential of the facultative amoebal parasite Acanthamoeba. This is the first study of its asparagine-linked glycans, for which we applied biochemical tools and various approaches of mass spectrometry. An initial glycomic screen of eight strains from five genotypes of this human pathogen suggested, in addition to the common eukaryotic oligomannose structures, the presence of pentose and deoxyhexose residues on their N-glycans. A more detailed analysis was performed on the N-glycans of a genotype T11 strain (4RE); fractionation by HPLC and tandem mass spectrometric analyses indicated the presence of a novel mannosylfucosyl modification of the reducing terminal core as well as phosphorylation of mannose residues, methylation of hexose and various forms of pentosylation. The largest N-glycan in the 4RE strain contained two N-acetylhexosamine, thirteen hexose, one fucose, one methyl, and two pentose residues; however, in this and most other strains analyzed, glycans with compositions of Hex(8-9)HexNAc(2)Pnt(0-1) tended to dominate in terms of abundance. Although no correlation between pathogenicity and N-glycan structure can be proposed, highly unusual structures in this facultative parasite can be found which are potential virulence factors or therapeutic targets.

AB - Glycans play key roles in host-pathogen interactions; thus, knowing the N-glycomic repertoire of a pathogen can be helpful in deciphering its methods of establishing and sustaining a disease. Therefore, we sought to elucidate the glycomic potential of the facultative amoebal parasite Acanthamoeba. This is the first study of its asparagine-linked glycans, for which we applied biochemical tools and various approaches of mass spectrometry. An initial glycomic screen of eight strains from five genotypes of this human pathogen suggested, in addition to the common eukaryotic oligomannose structures, the presence of pentose and deoxyhexose residues on their N-glycans. A more detailed analysis was performed on the N-glycans of a genotype T11 strain (4RE); fractionation by HPLC and tandem mass spectrometric analyses indicated the presence of a novel mannosylfucosyl modification of the reducing terminal core as well as phosphorylation of mannose residues, methylation of hexose and various forms of pentosylation. The largest N-glycan in the 4RE strain contained two N-acetylhexosamine, thirteen hexose, one fucose, one methyl, and two pentose residues; however, in this and most other strains analyzed, glycans with compositions of Hex(8-9)HexNAc(2)Pnt(0-1) tended to dominate in terms of abundance. Although no correlation between pathogenicity and N-glycan structure can be proposed, highly unusual structures in this facultative parasite can be found which are potential virulence factors or therapeutic targets.

KW - Animals

KW - Humans

KW - Carbohydrate Conformation

KW - Polysaccharides/chemistry/metabolism

KW - Mass Spectrometry/methods

KW - Acanthamoeba/chemistry/metabolism

KW - Glycomics

KW - Animals

KW - Humans

KW - Carbohydrate Conformation

KW - Polysaccharides/chemistry/metabolism

KW - Mass Spectrometry/methods

KW - Acanthamoeba/chemistry/metabolism

KW - Glycomics

M3 - SCORING: Journal article

VL - 287

SP - 43191

EP - 43204

JO - J BIOL CHEM

JF - J BIOL CHEM

SN - 0021-9258

IS - 52

M1 - 52

ER -