#METABOLOMICS WORKBENCH taq5016_20220518_084003 DATATRACK_ID:3256 STUDY_ID:ST002181 ANALYSIS_ID:AN003572 PROJECT_ID:PR001388
VERSION             	1
CREATED_ON             	May 30, 2022, 7:54 am
#PROJECT
PR:PROJECT_TITLE                 	Piperaquine-resistant PfCRT mutations differentially impact drug transport,
PR:PROJECT_TITLE                 	hemoglobin catabolism and parasite physiology in Plasmodium falciparum asexual
PR:PROJECT_TITLE                 	blood stages.
PR:PROJECT_SUMMARY               	The emergence of multidrug resistance in Plasmodium falciparum parasites
PR:PROJECT_SUMMARY               	presents a significant obstacle to the malaria elimination agenda. Resistance to
PR:PROJECT_SUMMARY               	piperaquine (PPQ), an important first-line partner drug, has spread across
PR:PROJECT_SUMMARY               	Southeast Asia where it has contributed to widespread treatment failures. The
PR:PROJECT_SUMMARY               	genetic cause of resistance to PPQ is attributable to a novel set of amino acid
PR:PROJECT_SUMMARY               	substitutions in the P. falciparum chloroquine resistance transporter (PfCRT).
PR:PROJECT_SUMMARY               	In this study, we used magnetically-purified trophozoite extracts from seven
PR:PROJECT_SUMMARY               	different lines comprising five genetically-modified and two field isolates.
PR:PROJECT_SUMMARY               	Three independent extractions with triplicate technical repeats were run by mass
PR:PROJECT_SUMMARY               	spectrometry on positive and negative modes and spectral peak raw data analyzed
PR:PROJECT_SUMMARY               	to obtain the fold change difference between the samples and parental control,
PR:PROJECT_SUMMARY               	Dd2Dd2crt. We show that PPQ-resistant, PfCRT mutant asexual blood stage
PR:PROJECT_SUMMARY               	parasites accumulate higher levels of hemoglobin-derived peptides than do their
PR:PROJECT_SUMMARY               	PPQ-sensitive counterparts.
PR:INSTITUTE                     	The Pennsylvania State University
PR:DEPARTMENT                    	Department of Biochemistry and Molecular Biology
PR:LAST_NAME                     	Llinas
PR:FIRST_NAME                    	Manuel
PR:ADDRESS                       	W126 Millenium Science Complex, University Park, PA 16802
PR:EMAIL                         	manuel@psu.edu
PR:PHONE                         	814-867-3527
#STUDY
ST:STUDY_TITLE                   	Piperaquine-resistant PfCRT mutations differentially impact drug transport,
ST:STUDY_TITLE                   	hemoglobin catabolism and parasite physiology in Plasmodium falciparum asexual
ST:STUDY_TITLE                   	blood stages.
ST:STUDY_SUMMARY                 	The emergence of multidrug resistance in Plasmodium falciparum parasites
ST:STUDY_SUMMARY                 	presents a significant obstacle to the malaria elimination agenda. Resistance to
ST:STUDY_SUMMARY                 	piperaquine (PPQ), an important first-line partner drug, has spread across
ST:STUDY_SUMMARY                 	Southeast Asia where it has contributed to widespread treatment failures. The
ST:STUDY_SUMMARY                 	genetic cause of resistance to PPQ is attributable to a novel set of amino acid
ST:STUDY_SUMMARY                 	substitutions in the P. falciparum chloroquine resistance transporter (PfCRT).
ST:STUDY_SUMMARY                 	In this study, we used magnetically-purified trophozoite extracts from seven
ST:STUDY_SUMMARY                 	different lines comprising five genetically-modified and two field isolates.
ST:STUDY_SUMMARY                 	Three independent extractions with triplicate technical repeats were run by mass
ST:STUDY_SUMMARY                 	spectrometry on positive and negative modes and spectral peak raw data analyzed
ST:STUDY_SUMMARY                 	to obtain the fold change difference between the samples and parental control,
ST:STUDY_SUMMARY                 	Dd2Dd2crt. We show that PPQ-resistant, PfCRT mutant asexual blood stage
ST:STUDY_SUMMARY                 	parasites accumulate higher levels of hemoglobin-derived peptides than do their
ST:STUDY_SUMMARY                 	PPQ-sensitive counterparts.
ST:INSTITUTE                     	The Pennsylvania State University
ST:DEPARTMENT                    	Department of Biochemistry and Molecular Biology
ST:LAST_NAME                     	Llinas
ST:FIRST_NAME                    	Manuel
ST:ADDRESS                       	W126 Millenium Science Complex, University Park, PA 16802
ST:EMAIL                         	manuel@psu.edu
ST:PHONE                         	814-867-3527
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Plasmodium falciparum
SU:TAXONOMY_ID                   	5833
SU:GENOTYPE_STRAIN               	DD2B2
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11082018-1	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11082018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11082018-2	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11082018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11082018-3	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11082018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11102018-1	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11102018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11102018-2	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11102018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11102018-3	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11102018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11122018-1	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11122018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11122018-2	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11122018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2B2-11122018-3	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2B2-11122018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10012018-1	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10012018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10012018-2	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10012018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10012018-3	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10012018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10062018-1	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10062018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10062018-2	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10062018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10062018-3	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10062018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10272018-1	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10272018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10272018-2	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10272018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crt-10272018-3	PfCRT Haplotype:Dd2	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crt-10272018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-06282018-1	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-06282018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-06282018-2	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-06282018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-06282018-3	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-06282018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-06302018-1	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-06302018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-06302018-2	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-06302018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-06302018-3	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-06302018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-07042018-1	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-07042018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-07042018-2	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-07042018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtF145I-07042018-3	PfCRT Haplotype:Dd2+F145I	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtF145I-07042018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-07212018-1	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-07212018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-07212018-2	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-07212018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-07212018-3	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-07212018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-07282018-1	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-07282018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-07282018-2	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-07282018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-07282018-3	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-07282018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-08022018-1	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-08022018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-08022018-2	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-08022018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtG353V-08022018-3	PfCRT Haplotype:Dd2+G353V	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtG353V-08022018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10092018-1	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10092018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10092018-2	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10092018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10092018-3	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10092018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10132018-1	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10132018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10132018-2	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10132018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10132018-3	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10132018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10152018-1	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10152018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10152018-2	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10152018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2Dd2crtM343L-10152018-3	PfCRT Haplotype:Dd2+M343L	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2Dd2crtM343L-10152018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10242018-1	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10242018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10242018-2	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10242018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10242018-3	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10242018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10292018-1	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10292018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10292018-2	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10292018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10292018-3	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10292018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10312018-1	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10312018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10312018-2	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10312018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	Dd2GC03crt-10312018-3	PfCRT Haplotype:3D7	Strain=Dd2; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=Dd2GC03crt-10312018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11132018-1	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11132018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11132018-2	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11132018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11132018-3	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11132018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11152018-1	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11152018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11152018-2	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11152018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11152018-3	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11152018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11172018-1	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11172018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11172018-2	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11172018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF12-11172018-3	PfCRT Haplotype:Dd2+H97Y	Strain=RF12; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF12-11172018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-07232018-1	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-07232018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-07232018-2	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-07232018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-07232018-3	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-07232018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-07312018-1	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-07312018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-07312018-2	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-07312018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-07312018-3	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-07312018-3.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-09102018-1	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-09102018-1.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-09102018-2	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-09102018-2.mzML
SUBJECT_SAMPLE_FACTORS           	-	RF7-09102018-3	PfCRT Haplotype:Dd2+M343L	Strain=RF7; Ion Mode=Negative/Positive; Parasite Stage=MACS purified Troph; RAW_FILE_NAME=RF7-09102018-3.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Mycoplasma-free parasites were sorbitol-synchronized in each generation for at
CO:COLLECTION_SUMMARY            	least two generations followed by magnetic enrichment of 32 hr post-invasion
CO:COLLECTION_SUMMARY            	trophozoites using MACS CS columns on a SuperMACS™ II Separator (Miltenyi
CO:COLLECTION_SUMMARY            	Biotec, Inc.) to remove uninfected RBCs. Trophozoite counts were determined on a
CO:COLLECTION_SUMMARY            	hemocytometer. Parasites were lysed in 1 mL of 90% cold methanol, containing 0.5
CO:COLLECTION_SUMMARY            	μM of the internal standard [13C4, 15N1]-Aspartate (Cambridge Isotope) to
CO:COLLECTION_SUMMARY            	correct for technical variations arising from sample processing. Samples were
CO:COLLECTION_SUMMARY            	vortexed to disrupt cell pellets and to generate uniform homogenates, which were
CO:COLLECTION_SUMMARY            	centrifuged (13,000´ g for 10 min) and the supernatants then harvested.
CO:COLLECTION_SUMMARY            	Supernatants were dried under nitrogen prior to resuspension in HPLC-grade water
CO:COLLECTION_SUMMARY            	for LC-MS analysis.
CO:SAMPLE_TYPE                   	Plasmodium cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Cells were all untreated, but had different genetic backgrounds.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Supernatants were dried under nitrogen prior to resuspension in HPLC-grade water
SP:SAMPLEPREP_SUMMARY            	for LC-MS analysis.
SP:PROCESSING_METHOD             	Lysis
SP:PROCESSING_STORAGE_CONDITIONS 	4℃
SP:EXTRACTION_METHOD             	See SA Cobbold et al JBC 2013
SP:EXTRACT_CLEANUP               	Clarification via centrifugation followed by nitrogen drying
SP:EXTRACT_STORAGE               	-80℃
SP:SAMPLE_RESUSPENSION           	HPLC-grade water spiked with chlorpropamide
SP:SAMPLE_SPIKING                	Internal Standard 13C4, 15N1-Aspartate used for sample preparation normalizer
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Samples (5ul) were separated by reverse phase HPLC using a Prominence 20 UFLCXR
CH:CHROMATOGRAPHY_SUMMARY        	system (Shimadzu, Columbia, MD) with a Waters (Milford, MA) BEH C18 column
CH:CHROMATOGRAPHY_SUMMARY        	(100mm x 2.1mm 1.7 um particle size) maintained at 55C and a 20 minute aqueous
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile gradient, at a flow rate of 250 ul/min. Solvent A was HPLC grade
CH:CHROMATOGRAPHY_SUMMARY        	water with 0.1% formic acid and Solvent B was HPLC grade acetonitrile with 0.1%
CH:CHROMATOGRAPHY_SUMMARY        	formic acid. The initial condition were 97% A and 3 % B, increasing to 45% B at
CH:CHROMATOGRAPHY_SUMMARY        	10 min, 75% B at 12 min where it was held at 75% B until 17.5 min before
CH:CHROMATOGRAPHY_SUMMARY        	returning to the initial conditions.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Prominence 20 UFLCXR
CH:COLUMN_NAME                   	Waters Acquity BEH C18 (100 x 2mm, 1.7um)
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	ABI Sciex 5600 TripleTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The eluate was delivered into a 5600 (QTOF) TripleTOF using a Duospray™ ion
MS:MS_COMMENTS                   	source (all Sciex, Framingham, MA).The capillary voltage was set at 5.5 kV in
MS:MS_COMMENTS                   	positive/negative ion mode with a declustering potential of 80V. The mass
MS:MS_COMMENTS                   	spectrometer was operated with a 100 ms TOF scan from 50 to 1000 m/z, and 16
MS:MS_COMMENTS                   	MS/MS product ion scans (100 ms) per duty cycle using a collision energy of 50V
MS:MS_COMMENTS                   	with a 20V spread.
MS:MS_RESULTS_FILE               	ST002181_AN003572_Results.txt	UNITS:peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END