Protein engineering as a strategy to avoid formation of amyloid fibrils.
The activation domain of human procarboxypeptidase A2 (ADA2h) aggregates following thermal or chemical denaturation at acidic pH. The aggregated material contains well-defined ordered structures with all the characteristics of the fibrils associated with amyloidotic diseases. Variants of ADA2h containing a series of mutations designed to increase the local stability of each of the two helical regions of the protein have been found to have a substantially reduced propensity to form fibrils. This arises from a reduced tendency of the denatured species to aggregate rather than from a change in the overall stability of the native state. The reduction in aggregation propensity may result from an increase in the stability of local relative to longer range interactions within the polypeptide chain. These findings show that the intrinsic ability of a protein to form amyloid can be altered substantially by protein engineering methods without perturbing significantly its overall stability or activity. This suggests new strategies for combating diseases associated with the formation of aggregated proteins and for the design of novel protein or peptide therapeutics.
Villegas
V
V
Departament de Bioquímica i Biologia Molecular, i Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Zurdo
J
J
Filimonov
V V
VV
Avilés
F X
FX
Dobson
C M
CM
Serrano
L
L
D000595:Amino Acid Sequence; D000682:Amyloid / Q000096:biosynthesis* / Q000737:chemistry; D002268:Carboxypeptidases / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D043422:Carboxypeptidases A; D002942:Circular Dichroism; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011489:Protein Denaturation; D015202:Protein Engineering; D017434:Protein Structure, Tertiary
D000682:Amyloid; D004792:Enzyme Precursors; D002268:Carboxypeptidases; D043422:Carboxypeptidases A