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. 2019 May 31;9:8120. doi: 10.1038/s41598-019-44542-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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PfpTKL SAM domain interacts specifically with PfSERA5. (a) PfSERA5 is the main binding partner of the PfpTKL SAM. P. falciparum proteins interacting with the PfpTKL SAM + RVxF1 region were identified by yeast two-hybrid (Y2H) screening. The percentage occurrence of each interacting protein is plotted for clones in frame with GAL4AD. All the hits returned by the screening are available in Supplementary Table S5. (b) PfSERA5 interacts strongly and specifically with the PfpTKL SAM in yeast. The chart shows the results of the mating run between Y2Hgold cells transformed with pGADT7 (either empty (Ø) or cloned with the longest PfSERA5 cDNA sequenced during Y2H screening) and Y187 cells transformed with pGBKT7 (empty (Ø), encoding laminin as a negative interaction control or SAM + RVxF-1 or SAM only). Diploids were either streaked on TDO/A medium (left panel) or QDO/A medium (right panel). (c) PfSERA5 interacts directly with the PfpTKL SAM in vitro. His₆-tagged PfSERA5 (His₆-SERA5) was incubated with pTKL_SAM-GST, pTKL_RVxF1-GST, PfeIf2γ-GST, PfPP1c-GST or GST (the last three = negative controls). After washes, proteins were eluted in Laemmli buffer, separated by SDS-PAGE and detected by western blot (WB, right panel). Inputs were also detected by western blot (left and middle panels). His₆-tagged PfSERA5 binds directly to pTKL_SAM-GST (first lane on right). Uncropped blots are available in the Supplementary Information.