Plasmodium falciparum infected red blood cells (iRBC) can adhere to endothelial cells of the blood-vessel wall, causing severe side effects. This adhesion is mediated by parasite proteins from the P. falciparum erythrocyte membrane protein 1 (PfEMP1) family, which localises to distinct protrusions on the iRBC membrane known as knobs. PfEMP1 members are transmembrane proteins with highly divergent extracellular regions that bind a range of endothelial cell receptors, such as ICAM-1 and CD36, and a predominantly flexible intracellular region, known as the acidic terminal segment (ATS). NMR studies have shown that ATS consists of three distinct flexible regions separated by a small helical core. Here we show that the N-terminal region of ATS interacts with cytoskeletal components, such as α and β spectrin, while the C-terminal region binds to members of the P. falciparum PHIST family PFI1780w and PFE1605. A combination of NMR, X-ray crystallography, mutagenesis, binding studies and computational methods such as Rosetta structure prediction and molecular dynamics have been used to build a larger interaction model for the ATS within the structure of the knob.