Lipid-derived aldehydes accelerate light chain amyloid and amorphous aggregation

Biochemistry. 2008 Jul 22;47(29):7695-705. doi: 10.1021/bi800333s. Epub 2008 Jun 26.

Abstract

Antibody light chain (LC) aggregation in vivo leads to the systemic deposition of Ig light chain domains in the form of either amyloid fibrils (AL-amyloidosis) or amorphous deposits, light-chain deposition disease (LCDD), in mainly cardiac or renal tissue and is a pathological condition that is often fatal. Molecular factors that may contribute to the propensity of LCs to aggregate in vivo, such as the protein primary structure or local environment, are intensive areas of study. Herein, we show that the aggregation of a human antibody kappa-(kappa-MJM) and lambda-(lambda-L155) light chain (1 mg/mL) can be accelerated in vitro when they are incubated under physiologically relevant conditions, PBS, pH 7.4 and 37 degrees C, in the presence of a panel of biologically relevant lipid-derived aldehydes, 4-hydroxynonenal (4-HNE), malondialdehyde (MDA), glyoxal (GLY), atheronal-A (KA), and atheronal-B (ALD). Thioflavin-T (ThT) and Congo Red (CR) binding assays coupled with turbidity studies reveal that this aldehyde-induced aggregation can be associated with alteration of protein secondary structure to an increased beta-sheet conformation. We observed that the nature of the conformational change is primarily dependent upon the lipidic aldehyde studied, not the protein sequence. Thus, the cholesterol 5,6-secosterols, KA and ALD, cause an amorphous-type aggregation which is ThT and CR negative for both the kappa-MJM and lambda-L155 light chains, whereas 4-HNE, MDA, and GLY induce aggregates that bind both ThT and CR. TEM analysis revealed that amyloid fibrils were formed during the 4-HNE-mediated aggregation of kappa-MJM and lambda-L155 light chains, whereas ALD-induced aggregates of these LCs where amorphous in nature. Kinetic profiles of LC aggregation reveal clear differences between the aldehydes, KA and ALD, causing a classic nucleated polymerization-type aggregation, with a lag phase (of approximately 150 h) followed by a growth phase that plateaus, whereas 4-HNE, MDA, and GLY trigger a seeded-type aggregation process that has no lag phase. In-depth studies of the 4-HNE-accelerated aggregation of kappa-MJM and lambda-L155 reveal a clear aldehyde concentration dependence and a process that can be inhibited by the naturally occurring osmolyte trimethylamine N-oxide (TMAO). Given these data, we feel our recently discovered paradigm of inflammatory aldehyde-induced protein misfolding may now extend to LC aggregation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehydes / chemistry*
  • Aldehydes / pharmacology
  • Benzothiazoles
  • Cholesterol / analogs & derivatives
  • Cholesterol / chemistry
  • Cholesterol / pharmacology
  • Congo Red / chemistry
  • Glyoxal / chemistry
  • Glyoxal / pharmacology
  • Humans
  • Immunoglobulin Light Chains / chemistry*
  • Immunoglobulin Light Chains / genetics
  • Immunoglobulin Light Chains / metabolism
  • Lipids / chemistry*
  • Malondialdehyde / chemistry
  • Malondialdehyde / pharmacology
  • Protein Denaturation / drug effects
  • Thiazoles / chemistry

Substances

  • Aldehydes
  • Benzothiazoles
  • Immunoglobulin Light Chains
  • Lipids
  • Thiazoles
  • atheronal-A
  • atheronal-B
  • thioflavin T
  • Congo Red
  • Malondialdehyde
  • Glyoxal
  • Cholesterol
  • 4-hydroxy-2-nonenal