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The Molecular Chaperone, α-Crystallin, Inhibits Amyloid Formation by Apolipoprotein C-II

2001; Elsevier BV; Volume: 276; Issue: 36 Linguagem: Inglês

10.1074/jbc.m105285200

1083-351X

Danny M. Hatters, Robyn A. Lindner, John A. Carver, Geoffrey J. Howlett,

Enzyme Structure and Function

Under lipid-free conditions, human apolipoprotein C-II (apoC-II) exists in an unfolded conformation that over several days forms amyloid ribbons. We examined the influence of the molecular chaperone, α-crystallin, on amyloid formation by apoC-II. Time-dependent changes in apoC-II turbidity (at 0.3 mg/ml) were suppressed potently by substoichiometric subunit concentrations of α-crystallin (1–10 μg/ml). α-Crystallin also inhibits time-dependent changes in the CD spectra, thioflavin T binding, and sedimentation coefficient of apoC-II. This contrasts with stoichiometric concentrations of α-crystallin required to suppress the amorphous aggregation of stressed proteins such as reduced α-lactalbumin. Two pieces of evidence suggest that α-crystallin directly interacts with amyloidogenic intermediates. First, sedimentation equilibrium and velocity experiments exclude high affinity interactions between α-crystallin and unstructured monomeric apoC-II. Second, the addition of α-crystallin does not lead to the accumulation of intermediate sized apoC-II species between monomer and large aggregates as indicated by gel filtration and sedimentation velocity experiments, suggesting that α-crystallin does not inhibit the relatively rapid fibril elongation upon nucleation. We propose that α-crystallin interacts stoichiometrically with partly structured amyloidogenic precursors, inhibiting amyloid formation at nucleation rather than the elongation phase. In doing so, α-crystallin forms transient complexes with apoC-II, in contrast to its chaperone behavior with stressed proteins.