Native state hydrogen exchange study of suppressor and pathogenic variants of transthyretin.
Transthyretin (TTR) is an amyloidogenic protein whose aggregation is responsible for numerous familial amyloid diseases, the exact phenotype being dependent on the sequence deposited. Many familial disease variants display decreased stability in vitro, and early onset pathology in vivo. Only subtle structural differences were observed upon crystallographic comparison of the disease-associated variants to the T119M interallelic trans-suppressor. Herein three human TTR single amino acid variant homotetramers including two familial amyloidotic polyneuropathy (FAP) causing variants (V30M and L55P), and a suppressor variant T119M (known to protect V30M carriers from disease by trans-suppression) were investigated in a residue-specific fashion by monitoring (2)H-(1)H exchange employing NMR spectroscopy. The measured protection factors for slowly exchanging amide hydrogen atoms reveal destabilization of the protein core in the FAP variants, the core consisting of strands A, B, E and G and the loop between strands A and B. The same core exhibits much slower exchange in the suppressor variant. Accelerated exchange rates were observed for residues at the subunit interfaces in L55P, but not in the T119M or V30M TTR. The correlation between destabilization of the TTR core strands and the tendency for amyloid formation supports the view that these strands are involved in amyloidogenicity, consistent with previous (2)H-(1)H exchange analysis of the WT-TTR amyloidogenic intermediate.
Liu
Kai
K
Department of Chemistry and Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, BCC260, La Jolla, CA 92037, USA.
Kelly
Jeffery W
JW
Wemmer
David E
DE
D019943:Amino Acid Substitution; D028227:Amyloid Neuropathies, Familial / Q000235:genetics* / Q000473:pathology; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics*; D014644:Genetic Variation; D006801:Humans; D006859:Hydrogen; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011228:Prealbumin / Q000737:chemistry / Q000235:genetics*; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary
D002352:Carrier Proteins; D011228:Prealbumin; D006859:Hydrogen