Mechanistic Insights into the Early Events in the Aggregation of Immunoglobulin Light Chains

Pinaki Misra, Luis M. Blancas-Mejia, Marina Ramirez-Alvarado

Research output: Contribution to journalArticle

Abstract

Little is known about the mechanism of amyloid assembly in immunoglobulin light chain (AL) amyloidosis, in contrast to other amyloid diseases. Early events in the aggregation pathway are especially important, as these soluble species could be cytotoxic intermediates playing a critical role in the initiation of the amyloid assembly. In this work, we discuss the mechanism of the early events in in vitro fibril formation of immunoglobulin light chain AL-09 and AL-12 (involved in cardiac amyloidosis) and its germline (control) protein κI O18/O8. Previous work from our laboratory showed that AL-12 adopts a canonical dimer conformation (like the germline protein), whereas AL-09 presents an altered dimer interface as a result of somatic mutations. Both AL-12 and AL-09 aggregate with similar rates and significantly faster than the germline protein. AL-09 is the only protein in this study that forms stable oligomeric intermediates during the early stages of the aggregation reaction with some structural rearrangements that increase the thioflavin T fluorescence but maintain the same number of monomers in solution. The presence of the restorative mutation AL-09 H87Y changes the kinetics and the aggregation pathway compared to AL-09. The single restorative mutation AL-12 R65S slightly delayed the overall rate of aggregation as compared to AL-12. Collectively, our study provides a comprehensive analysis of species formed during amyloid nucleation in AL amyloidosis, shows a strong dependence between the altered dimer conformation and the formation of stable oligomeric intermediates, and sheds light on the structural features of amyloidogenic intermediates associated with cellular toxicity.

Original languageEnglish (US)
Pages (from-to)3155-3168
Number of pages14
JournalBiochemistry
Volume58
Issue number29
DOIs
StatePublished - Jun 25 2019

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Immunoglobulin Light Chains
Amyloid
Amyloidosis
Agglomeration
Dimers
Mutation
Conformations
Proteins
Toxicity
Nucleation
Monomers
Fluorescence
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mechanistic Insights into the Early Events in the Aggregation of Immunoglobulin Light Chains. / Misra, Pinaki; Blancas-Mejia, Luis M.; Ramirez-Alvarado, Marina.

In: Biochemistry, Vol. 58, No. 29, 25.06.2019, p. 3155-3168.

Research output: Contribution to journalArticle

Misra, Pinaki ; Blancas-Mejia, Luis M. ; Ramirez-Alvarado, Marina. / Mechanistic Insights into the Early Events in the Aggregation of Immunoglobulin Light Chains. In: Biochemistry. 2019 ; Vol. 58, No. 29. pp. 3155-3168.
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