Rising, Anna
- Institutionen för husdjurens biovetenskaper, Sveriges lantbruksuniversitet
- Karolinska institutet
Sammanfattning
Amyloidogenesis is associated with more than 30 diseases, but the molecular mechanisms involved in cell toxicity and fibril formation remain largely unknown. The inherent tendency of amyloid-forming proteins to aggregate renders expression, purification, and experimental studies challenging. NT* is a solubility tag derived from a spider silk protein that was recently introduced for the production of several aggregation-prone peptides and proteins at high yields. Herein, we investigate whether fusion to NT* can prevent amyloid fibril formation and enable controlled aggregation for experimental studies. As an example of an amyloidogenic protein, we chose the de novo-designed polypeptide 17. The fusion protein NT*-17 was recombinantly expressed in Escherichia coli to produce high amounts of soluble and mostly monomeric protein. Structural analysis showed that 17 is kept in a largely unstructured conformation in fusion with NT*. After proteolytic release, 17 adopts a -sheet conformation in a pH- and salt-dependent manner and assembles into amyloid-like fibrils. The ability of NT* to prevent premature aggregation and to enable structural studies of prefibrillar states may facilitate investigation of proteins involved in amyloid diseases.
Nyckelord
amyloid disease; fibril formation; model protein; protein assembly; protein domain
Publicerad i
FEBS Journal
2018, volym: 285, nummer: 10, sidor: 1873-1885
Utgivare: WILEY
SLU författare
Johansson, Jan
- Karolinska institutet
UKÄ forskningsämne
Biokemi
Molekylärbiologi
Publikationens identifierare
- DOI: https://doi.org/10.1111/febs.14451
Permanent länk till denna sida (URI)
https://res.slu.se/id/publ/95808