Binding of amyloidogenic transthyretin to the plasma membrane alters membrane fluidity and induces neurotoxicity.
Transthyretin (TTR) can deposit as amyloid in the peripheral nervous system and induce a peripheral neuropathy. We examined the mechanism of TTR amyloid neurotoxicity on SH-SY5Y neuroblastoma cells. Wild-type (WT) TTR and two amyloidogenic mutants (V30M and L55P) were expressed in Escherichia coli. Incubation (aging) of WT TTR at 37 degrees C for 1 week caused no significant aggregation. However, there was a significant increase in the extent of amyloid fibril formation after the amyloidogenic mutants had been aged. L55P TTR aggregated more readily than V30M TTR. Both amyloidogenic mutants were neurotoxic after aging. The order of neurotoxicity was as follows: L55P > V30M > WT. As binding of amyloid proteins to the plasma membrane may cause cytotoxicity, we studied the binding of TTR to a plasma membrane-enriched preparation from SH-SY5Y cells by surface plasmon resonance. All three forms bound to the plasma membrane through electrostatic interactions. The binding of the amyloidogenic mutants was increased by aging. The amount of binding correlated closely with the amount of aggregation and with the cytotoxicity of each form. As membrane fluidity can influence cell viability, we also examined the effect of TTR on membrane fluidity using a fluorescence anisotropy method. Binding of the amyloidogenic TTR mutants increased membrane fluidity, and once again, the order of potency was as follows: L55P > V30M > WT. These results demonstrate that TTR can bind to the plasma membrane and cause a change in membrane fluidity. Altered membrane fluidity may be the cause of the neurotoxicity.
Hou
Xu
X
Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3800, Australia.
Richardson
Samantha J
SJ
Aguilar
Marie-Isabel
MI
Small
David H
DH
D019943:Amino Acid Substitution; D001665:Binding Sites; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D008560:Membrane Fluidity / Q000502:physiology*; D016297:Mutagenesis, Site-Directed; D009447:Neuroblastoma; D009498:Neurotoxins / Q000633:toxicity*; D010957:Plasmids; D011228:Prealbumin / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D009498:Neurotoxins; D011228:Prealbumin; D011994:Recombinant Proteins