Aggregation of misfolded proteins can be a selective process dependent upon peptide composition

Micha I. Milewski, John E. Mickle, John K. Forrest, Bruce A. Stanton, Garry R Cutting

Research output: Contribution to journalArticle

Abstract

Intracellular aggregation of misfolded proteins is observed in a number of human diseases, in particular, neurologic disorders in which expanded tracts of polyglutamine residues play a central role. A variety of other proteins are prone to aggregation when mutated, indicating that this process is a common pathologic mechanism for inherited disorders. However, little is known about the relationship between the sequence of aggregating peptides and the specificity of intracellular accumulation. Here we demonstrate that substitution of two residues eliminates aggregation of a 111-amino acid peptide derived from the C-terminal portion of the cystic fibrosis transmembrane conductance regulator (CFTR). We also show that fusion to a reporter protein considerably alters the subcellular distribution of aggregating peptide. When fused to green fluorescent protein, the peptide containing amino acids 1370-1480 of CFTR accumulates in large perinuclear or nuclear aggregates. The same CFTR fragment devoid of green fluorescent protein localizes predominantly to discrete accumulations associated with mitochondria. Importantly, both types of accumulation are dependent on the presence of the same two amino acids within the CFTR sequence. Co-expression studies show that both CFTR-derived proteins can co-localize in large cytoplasmic/nuclear aggregates. However, neither CFTR construct accumulates in intracellular inclusions formed by N-terminal fragment of huntingtin. In addition to unique accumulation patterns, each aggregating peptide shows differences in association with chaperone proteins. Thus, our results indicate that the process of intracellular aggregation can be a selective process determined by the composition of the aggregating peptides.

Original languageEnglish (US)
Pages (from-to)34462-34470
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number37
DOIs
StatePublished - Sep 13 2002

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Cystic Fibrosis Transmembrane Conductance Regulator
Agglomeration
Peptides
Chemical analysis
Proteins
Green Fluorescent Proteins
Amino Acids
Mitochondria
Nervous System Diseases
Substitution reactions
Fusion reactions
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Aggregation of misfolded proteins can be a selective process dependent upon peptide composition. / Milewski, Micha I.; Mickle, John E.; Forrest, John K.; Stanton, Bruce A.; Cutting, Garry R.

In: Journal of Biological Chemistry, Vol. 277, No. 37, 13.09.2002, p. 34462-34470.

Research output: Contribution to journalArticle

Milewski, Micha I. ; Mickle, John E. ; Forrest, John K. ; Stanton, Bruce A. ; Cutting, Garry R. / Aggregation of misfolded proteins can be a selective process dependent upon peptide composition. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 37. pp. 34462-34470.
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