Sequential truncation of the spacer region caused trapping of the Nluc-mDH-FL constructs in the ER and PV of the parasite.(A) Representative IFA images (n = 3 independent replicates) of HSP101-HAglmS parasites expressing Hyp1/STEVOR/KAHRP-Nluc-mDH-FLproteins with various spacer lengths (see diagrams). Parasite cells were probed with anti-EXP2 and anti-HA antibodies to visualise the PTEX components within the cell. Anti-PfERC antibody was used to visualise parasite's ER. The Nluc-mDH-FL proteins were either probed with anti-Nluc oranti-FLAG antibodies. Scale bars, 5 μm. DIC, Differential Interference Contrast. DAPI (40,6-diamidino-2-phenylindole; blue) was used to stain parasite nuclei. (B) Quantification of export of the spacer truncation Nluc-mDH-FL constructs presented in (A). The following numbers of cells were used for the analysis. Hyp1 constructs: n = 25 (51aa), 25 (13aa), 28 (3aa) cells; STEVOR constructs: n = 53 (49aa), 32 (13aa), 23 (13aa) cells; KAHRP constructs: n = 26 (49aa), 23 (13aa), 30 (3aa) cells. Box and whisker plot represents 25th–75th and 10th–90th percentiles, respectively. Statistical significance was determined using one-way ANOVA with Brown-Forsythe test followed by Tukey’s multiple comparisons test. ****, p-value<0.0001; **, p-value<0.01; *,p-value = 0.0159). Gabriela M, Barnes CBG, Leong D, Sleebs BE, Schneider MP, Littler DR, Crabb BS, de Koning-Ward TF, Gilson PR. Sequence elements within the PEXEL motif and its downstream region modulate PTEX-dependent protein export in Plasmodium falciparum. Traffic. 2024 Jan;25(1):e12922. doi: 10.1111/tra.12922. Epub 2023 Nov 5. PMID: 37926971
Other associated proteins
PFID | Formal Annotation |
---|---|
PF3D7_0101800 | stevor PIR protein |
PF3D7_0202000 | knob-associated histidine-rich protein |