posted on 2021-02-17, 16:05authored byYibo Wang, Bo Zhang, Cong Lin, Ying Liu, Min Yang, Yinghua Peng, Xiaohui Wang
Charged
residues are frequently found in the transmembrane segments
of membrane proteins, which reside in the hydrophobic bilayer environment.
Charged residues are critical for the function of membrane protein.
However, studies of their role in protein oligomerization are limited.
By taking the fifth transmembrane domain (TMD5) of latent membrane
protein 1 from the Epstein–Barr virus as a prototype model, in silico simulations and wet-lab experiments were performed
to investigate how the charged states affect transmembrane domain
oligomerization. Molecular dynamics (MD) simulations showed that the
D150-protonated TMD5 trimer was stable, whereas unprotonated D150
created bends in the helices which distort the trimeric structure.
D150 was mutated to asparagine to mimic the protonated D150 in TMD5,
and the MD simulations of different D150N TMD5 trimers supported that
the protonation state of D150 was critical for the trimerization of
TMD5. In silico mutations found that D150N TMD5 preferred
to interact with TMD5 to form the heterotrimer (1 D150N TMD5:2 protonated
TMD5s) rather than the heterotrimer (2 D150N TMD5s:1 protonated TMD5).
D150R TMD5 interacted with TMD5 to form the heterotrimer (1 D150R
TMD5:2 protonated TMD5). These in silico results
imply that D150N TMD5 and D150R TMD5 peptides may be probes for disrupting
TMD5 trimerization, which was supported by the dominant-negative ToxR
assay in bacterial membranes. In all, this study elucidates the role
of charged residues at the membrane milieu in membrane protein oligomerization
and provides insight into the development of oligomerization-regulating
peptides for modulating transmembrane domain lateral interactions.