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. 2007 May 30;2(5):e477. doi: 10.1371/journal.pone.0000477

Figure 5. The DBL domain of Pf332 binds to human RBC.

Figure 5

A: The protein Pf332 consists of a potentially extracellular region encoded by exon I and is composed of a DBL- and nDBL-domain followed by a predicted transmembrane domain. The region of the molecule originally described as Pf332 is encoded by a second exon and contains a Glutamic-acid rich part, which includes a Valin-rich area as well as the region EB200 build of numerous repeats of a XXXEEXXEEXX motif (X = hydrophobic aa). B: The Pf332-DBL- and Pf332nDBL-region were expressed as two individual GST-fusion proteins in E.coli. Binding assays, in which human RBC were incubated with the DBL-, nDBL- and an unrelated control protein were carried out. Binding of the proteins was visualized by subjecting the RBC to immunoblotting, where the bound protein was detected with an α-GST antibody. The DBL- but not the nDBL-domain or the unrelated control protein was able to bind to human RBC. C: Transient transfected CHO cells expressing either the DBL- or nDBL-domain on their surface were tested for their ability to bind to human RBC. CHO cells were stained red with PHK26 and incubated with RBC stained green with PKH67. CHO-cells expressing the DBL-domain avidly bound RBC, lower panel, while CHO cells expressing the nDBL-region or MOCK-transfected cells did not show any binding towards RBC (middle and upper panel).