title: Sugar nucleotide quantification by liquid chromatography tandem mass spectrometry reveals a distinct profile inPlasmodium falciparumsexual stage parasites.
abstract: The obligate intracellular lifestyle ofPlasmodium falciparumand the difficulties in obtaining sufficient amounts of biological material have hampered the study of specific metabolic pathways in the malaria parasite. Thus, for example, the pools of sugar nucleotides required to fuel glycosylation reactions have never been studied in-depth in well-synchronized asexual parasites or in other stages of its life cycle. These metabolites are of critical importance, especially considering the renewed interest in the presence ofN-,O-, and other glycans in key parasite proteins. In this work, we adapted a liquid chromatography tandem mass spectrometry (LC-MS/MS) method based on the use of porous graphitic carbon (PGC) columns and MS-friendly solvents to quantify sugar nucleotides in the malaria parasite. We report the thorough quantification of the pools of these metabolites throughout the intraerythrocytic cycle ofP. falciparumThe sensitivity of the method enabled, for the first time, the targeted analysis of these glycosylation precursors in gametocytes, the parasite sexual stages that are transmissible to the mosquito vector.
authors.affiliation: ISGlobal, Barcelona Ctr. Int. Health Res. (CRESIB), Hospital Clínic - Universitat de Barcelona, Barcelona, Spain.
authors.forename: Borja
authors.initials: B
authors.lastname: López-Gutiérrez
authors.affiliation: Department of Infectious Diseases & Pathology, The University of Florida Emerging Pathogens Institute, Gainesville, FL 32611, U.S.A.
authors.forename: Rhoel R
authors.initials: RR
authors.lastname: Dinglasan
authors.affiliation: ISGlobal, Barcelona Ctr. Int. Health Res. (CRESIB), Hospital Clínic - Universitat de Barcelona, Barcelona, Spain luis.izquierdo@isglobal.org.
authors.forename: Luis
authors.initials: L
authors.lastname: Izquierdo
mesh_terms: D002853:Chromatography, Liquid; D004912:Erythrocytes / Q000469:parasitology; D005718:Gametogenesis / Q000502:physiology; D006154:Guanosine Diphosphate Fucose / Q000032:analysis / Q000378:metabolism*; D006155:Guanosine Diphosphate Mannose / Q000032:analysis / Q000378:metabolism*; D006156:Guanosine Diphosphate Sugars / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008018:Life Cycle Stages / Q000502:physiology; D010963:Plasmodium falciparum / Q000254:growth & development / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014531:Uridine Diphosphate Galactose / Q000032:analysis / Q000378:metabolism*; D014532:Uridine Diphosphate Glucose / Q000032:analysis / Q000378:metabolism*; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000032:analysis / Q000378:metabolism*
chemical_list: D006156:Guanosine Diphosphate Sugars; D006154:Guanosine Diphosphate Fucose; D014531:Uridine Diphosphate Galactose; D006155:Guanosine Diphosphate Mannose; C100085:guanosine diphosphate glucose; D014536:Uridine Diphosphate N-Acetylgalactosamine; D014532:Uridine Diphosphate Glucose