Nene, Vishvanath

Nyagwange, James

Pelle, Roger

Ternette, Nicola

Tijhaar, Edwin

Nene, Vishvanath M.

CGSpace (CGIAR)

Proteomics of Theileria parva sporozoitesJames Nyagwange 1, 3, Nicola Ternette 2, Edwin Tijhaar 3, Roger Pelle’ 4, Vish Nene 51, 4, 5 International Livestock Research Institute, P. O. Box 30709, Nairobi, Kenya2 The Jenner Institute, Nufﬁeld Department of Medicine, University of Oxford, UK3 Cell Biology and Immunology Group, Wageningen University, The NetherlandsIntroductionEast Coast fever (ECF) is a lymphoproliferative disease caused by the protozoan parasite Theileria parva. It kills about one million cattle annually in Africa. The sporozoite stage of this parasite, harbored in the salivary glands of the tick Rhipicephalus appendiculatus, invades and establishes infection in the bovine lymphocytes during tick feeding. However, little is known about the parasite molecules involved in this  infection process. It is therefore necessary to elucidate the protein composition of the sporozoites to identify novel targets for blocking invasion. Blocking this initial stage of invasion presents a promising vaccine strategy for the control of ECF. AimTo (i) establish the protein expression profile of T. parva sporozoites and (ii) identify proteins involved in host cell entry and invasionMethodResults OGs Organism name ORF locus tag Gene name AnnotationRhoptry neck proteinsOG5_142870 Plasmodium falciparum PF14_0495 PfRON2 Rhoptry neck protein 2Theileria parva TP01_0014 - Hypothetical proteinOG5_153587 Plasmodium falciparum MAL8P1.73 - Conserved Plasmodium protein, unknown functionTheileria parva TP01_1161 - Hypothetical proteinRhoptry ProteinsOG5_153563 Plasmodium falciparum MAL13P1.237 - Conserved Plasmodium protein, unknown functionTheileria parva TP02_0645 - Hypothetical proteinOG5_128020 Plasmodium falciparum PFI1570c PfM18AAP M18 aspartyl aminopeptidaseTheileria parva TP01_1150 - Aspartyl aminopeptidaseOG5_145111 Plasmodium falciparum PFI0295c - RNB-like protein, putativeTheileria parva TP01_0396 - Hypothetical proteinOG5_141731 Plasmodium falciparum MAL13P1.308 - Conserved Plasmodium protein, unknown functionTheileria parva TP01_0092 - Hypothetical proteinMicroneme ProteinsOG5_147452 Plasmodium falciparum PF11_0344 AMA1 Apical membrane antigen 1, AMA1Theileria parva TP01_0650 - Apical membrane antigen 1OG5_134280 Plasmodium falciparum PFL0805w - MAC/ Perforin, putativePlasmodium falciparum PF08_0052 - Perforin like protein 5Theileria parva TP01_0164 - Hypothetical proteinTheileria parva TP02_0166 - Hypothetical proteinOG5_171217 Plasmodium falciparum PFL0800c CelTOS CelTOS, putativeTheileria parva TP01_0232 - Hypothetical proteinOG5_135185 Plasmodium falciparum PFB0585w - Leu/Phe-tRNA protein transferase, putativeTheileria parva TP02_0627 - Leucyl/phenylalanyl-tRNA protein transferaseDense Granules/ ExonemesOG5_126706 Plasmodium falciparum MAL8P1.69 - 14-3-3 protein, putativeTheileria parva TP02_0607 - Hypothetical proteinOGs Organism name ORF locus tag Gene name AnnotationOG5_127135 Plasmodium falciparum PFB0210c HT1 Hexose transporter, PfHT1Theileria parva TP01_1069 - Monosaccharide transporterOG5_126744 Plasmodium falciparum PF11_0351 - Heat shock protein hsp70 homologuePlasmodium falciparum PF08_0054 - Heat shock 70 kDa proteinPlasmodium falciparum MAL13P1.540 - Heat shock protein 70 (hsp70), putativeTheileria parva TP01_0479 - Hypothetical proteinTheileria parva TP02_0148 - Heat shock protein 70Theileria parva TP02_0753 - Heat shock protein 70OG5_127061 Plasmodium falciparum PFD0305c - Vacuolar ATP synthase subunit bTheileria parva TP01_0389 - ATP synthase F1 subunit betaOG5_126628 Plasmodium falciparum PF14_0598 GAPDH Glyceraldehyde-3-phosphate dehydrogenaseTheileria parva TP02_0858 - Glyceraldehyde-3-phosphate dehydrogenaseOG5_143227 Plasmodium falciparum PF14_0065 -Conserved Plasmodium membrane protein, unknown functionTheileria parva TP01_0761 - Hypothetical proteinOG5_147452 Plasmodium falciparum PF11_0344 AMA1 Apical membrane antigen 1, AMA1Theileria parva TP01_0650 - Apical membrane antigen 1OG5_126595 Plasmodium falciparum PF11_0114 - Actin-like protein homolog, ALP1 homologPlasmodium falciparum PFL2215w ACT1 Actin IPlasmodium falciparum PF14_0124 - Actin IITheileria parva TP01_0789 - ActinTheileria parva TP02_0056 - ActinTheileria parva TP02_0903 - ActinORFlocus tagCoverage(%)# UniquePeptides DescriptionTMHMM GPI-SOM SignalIP Protter# TM Domains GPI-linkedSignal peptide # TM Domains GPI-linkedSignal peptide TP01_0012 42 6 14 kDa zinc-binding protein 1 1 0 1 0 0TP01_0438 24 34 Hypothetical protein 0 1 1 0 0 1TP01_0554 10 3 Hypothetical protein 0 1 0 0 0 0TP01_0655 5 3 E3 ubiquitin-protein ligase synoviolin 6 1 0 5 0 0TP01_0715 104Hypothetical protein 2 1 0 2 0 0TP01_0939 34 12 Hypothetical protein 1 1 1 1 0 1TP01_0972 16 4 Hypothetical protein 1 1 1 1 0 1TP01_1056 39 12 Merozoite Antigen 0 1 1 0 0 1TP01_1066 22 5 Hypothetical protein 5 1 0 5 0 0TP02_0412 11 5 Hypothetical protein 0 1 1 0 0 1TP02_0500 4 3 Sodium-dependent phosphate transporter 1-B 9 1 0 9 0 0TP02_0538 23 31 Hypothetical protein 1 1 1 1 0 1TP02_0553 30 6 Hypothetical protein 1 1 1 1 0 1TP02_0950 5 2 Hypothetical protein 1 1 1 1 0 1TP01_0876 16 1 Hypothetical protein 1 0 0 1 0 1TP03_0245 2 1 Hypothetical protein 9 1 0 3 0 0TP03_0287 68 53 Sporozoite P67 surface antigen 1 1 1 1 0 1TP03_0512 38 12 Tartrate-resistant acid phosphatase type 5 2 1 0 1 0 0TP04_0437 53 60 104 kDa microneme/rhoptry antigen 1 1 1 0 1 1TP04_0615 7 2Probable N- acetylglucosaminyl-phosphatidylinositol de-N- acetylase 0 1 1 0 0 1Table 1. Invasion organelle proteins. Proteins detected from whole cell lysates of T. parva sporozoites are aligned by orthology with P. falciparum sporozoites proteins into orthologous groups (OGs).Table 2. Proteins involved with calcium signaling. Proteins detected from whole cell lysates of T. parva sporozoites are aligned by orthology with P. falciparum sporozoites proteinsTable 3. Surface proteins. Proteins detected from whole cell lysates of T. parvasporozoites are aligned by orthology with P. falciparum sporozoites proteinsTable 4. Additional surface proteins. Proteins detected from whole cell lysates of T. parva sporozoites predicted to contain either GPI anchor, transmembrane domain or signal peptide.Total of 4474 proteins identified2006 = T. parva proteins2468 = tick proteinsAnalysis of 2006 T. parva proteins done by: TargetP – localizationPlasmoAP – apicoplastOrthoMCL – orthologyGPI-SOM – GPI anchorsTMHMM – transmembrane domains (TM)SignalIP – signal peptide (SP)Protter – GPI anchor, TM and SP Conclusion In total 2006 T. parva sporozoite proteins have been confirmed to be expressed representing approximately 50% of  the  total  predicted T. parva protein expression profile. Potential surface proteins and possible proteins involved in host entry and invasion have been identified using bioinformatic tools.OGs Organism name ORF locus tag Gene name Annotation OG5_147452 Plasmodium falciparum PF11_0344 AMA1 Apical membrane antigen 1, AMA1 Theileria parva TP01_0650 - Apical membrane antigen 1OG5_126800 Plasmodium falciparum PF14_0323 - CalmodulinTheileria parva TP02_0717 - CalmodulinOG5_129380 Plasmodium falciparum PFA_0345w PfCEN1 Centrin-1 Theileria parva TP01_0227 - CentrinOG5_152981 Plasmodium falciparum PF11_0411 - Conserved Plasmodium proteinTheileria parva TP02_0412 - Hypothetical proteinOG5_146625 Plasmodium falciparum PF11_0239 CDPK6 Calcium-dependent protein kinase, putative Plasmodium falciparum PF07_0072 PfCDPK4 Calcium-dependent protein kinase 4 Plasmodium falciparum PF13_0211 CDPK5 Calcium-dependent protein kinase, putative Theileria parva TP01_0983 - Calcium-dependent protein kinaseTheileria parva TP01_1073 - Calmodulin-domain protein kinaseTheileria parva TP02_0399 - Calcium-dependent protein kinaseOG5_133188 Plasmodium falciparum PF11_0395 - Guanylyl cyclase Theileria parva TP02_0848 - Guanylyl cyclaseOG5_141757 Plasmodium falciparum PFL2225w MTIP Myosin A tail domain interacting protein Theileria parva TP01_0513 - Myosin light chainOG5_126577 Plasmodium falciparum MAL13P1.148 - Myosin Theileria parva TP01_0774 - myosinOG5_126674 Plasmodium falciparum PFL0590c ATPase4 Non-SERCA-type Ca2+ -transporting P-ATPase Plasmodium falciparum PFA_0310c - Calcium-transporting ATPase, putative Theileria parva TP01_0720 - P-type ATPaseTheileria parva TP02_0524 - Calcium-transporting ATPaseOG5_127599 Plasmodium falciparum PF14_0346 PKG cGMP-dependent protein kinaseTheileria parva TP02_0378 - cAMP-dependent protein kinaseOG5_126635 Plasmodium falciparum PFL2250c PKB Rac-beta serine/threonine protein kinase, PfPKBTheileria parva TP02_0727 - cAMP-dependent protein kinase catalytic subunitThis poster is licensed for use under the Creative Commons Attribution 4.0 International Licence (May 2016)

Proteomics of Theileria parva sporozoites

https://cgspace.cgiar.org/bitstream/10568/75993/1/ecf_keystone_nyagwange.pdf

Proteomics of Theileria parva sporozoites

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