L55P transthyretin accelerates subunit exchange and leads to rapid formation of hybrid tetramers.
Transthyretin is a tetrameric protein associated with the commonest form of systemic amyloid disease. Using isotopically labeled proteins and mass spectrometry, we compared subunit exchange in wild-type transthyretin with that of the variant associated with the most aggressive form of the disease, L55P. Wild-type subunit exchange occurs via both monomers and dimers, whereas exchange via dimers is the dominant mechanism for the L55P variant. Because patients with the L55P mutation are heterozygous, expressing both proteins simultaneously, we also analyzed the subunit exchange reaction between wild-type and L55P tetramers. We found that hybrid tetramers containing two or three L55P subunits dominate in the early stages of the reaction. Surprisingly, we also found that, in the presence of L55P transthyretin, the rate of dissociation of wild-type transthyretin is increased. This implies interactions between the two proteins that accelerate the formation of hybrid tetramers, a result with important implications for transthyretin amyloidosis.
Keetch
Catherine A
CA
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
Bromley
Elizabeth H C
EH
McCammon
Margaret G
MG
Wang
Nan
N
Christodoulou
John
J
Robinson
Carol V
CV
D000682:Amyloid / Q000737:chemistry; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism; D006579:Heterozygote; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D008958:Models, Molecular; D009154:Mutation; D011228:Prealbumin / Q000737:chemistry* / Q000235:genetics*; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012984:Software; D013997:Time Factors
D000682:Amyloid; D011228:Prealbumin; D011994:Recombinant Proteins