Transthyretin slowly exchanges subunits under physiological conditions:: A convenient chromatographic method to study subunit exchange in oligomeric proteins
被引:88
作者:
Schneider, F
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机构:Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
Schneider, F
Hammarström, P
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机构:Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
Hammarström, P
Kelly, JW
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机构:Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
Kelly, JW
机构:
[1] Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
[2] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA
transthyretin;
amyloid;
familial amyloid polyneuropathy;
ion exchange chromatography;
D O I:
10.1110/ps.8901
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
Transthyretin (TTR) subunits were labeled with a charge-modifying tag to evaluate the possibility of subunit exchange between tetramers under physiological conditions. Starting with a mixture of two TTR homote-tramers, one having all subunits tagged at the N termini and the ether composed of untagged subunits, heterotetramer formation as a function of time and temperature was evaluated using ion exchange chromatography. The data indicate that the subunit exchange can occur under native conditions at physiological pH in vitro, albeit slowly. Wild-type TTR exchanges subunits on a timescale of days at 37 degreesC and on a timescale of hours at 4 degreesC. The familial amyloid polyneuropathy-associated variant V30M exchanges subunits at the same rate as wild-type TTR at 4 degreesC but slower and less efficiently at 37 degreesC. Small molecule tetramer stabilizers abolish TTR subunit exchange, supporting a dissociative mechanism.