Ligand binding promotes prion protein aggregation--role of the octapeptide repeats.
Aggregation of the normal cellular prion protein, PrP, is important in the pathogenesis of prion disease. PrP binds glycosaminoglycan (GAG) and divalent cations, such as Cu(2+) and Zn(2+). Here, we report our findings that GAG and Cu(2+) promote the aggregation of recombinant human PrP (rPrP). The normal cellular prion protein has five octapeptide repeats. In the presence of either GAG or Cu(2+), mutant rPrPs with eight or ten octapeptide repeats are more aggregation prone, exhibit faster kinetics and form larger aggregates than wild-type PrP. When the GAG-binding motif, KKRPK, is deleted the effect of GAG but not that of Cu(2+) is abolished. By contrast, when the Cu(2+)-binding motif, the octapeptide-repeat region, is deleted, neither GAG nor Cu(2+) is able to promote aggregation. Therefore, the octapeptide-repeat region is critical in the aggregation of rPrP, irrespective of the promoting ligand. Furthermore, aggregation of rPrP in the presence of GAG is blocked with anti-PrP mAbs, whereas none of the tested anti-PrP mAbs block Cu(2+)-promoted aggregation. However, a mAb that is specific for an epitope at the N-terminus enhances aggregation in the presence of either GAG or Cu(2+). Therefore, although binding of either GAG or Cu(2+) promotes the aggregation of rPrP, their aggregation processes are different, suggesting multiple pathways of rPrP aggregation.
Yu
Shuiliang
S
Department of Pathology, Case Western Reserve University, Cleveland, OH 44106-7288, USA.
Yin
Shaoman
S
Pham
Nancy
N
Wong
Poki
P
Kang
Shin-Chung
SC
Petersen
Robert B
RB
Li
Chaoyang
C
Sy
Man-Sun
MS
D020816:Amino Acid Motifs; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001665:Binding Sites; D002413:Cations, Divalent; D003300:Copper / Q000378:metabolism*; D000939:Epitopes; D006025:Glycosaminoglycans / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D009842:Oligopeptides / Q000378:metabolism; D011328:Prions / Q000378:metabolism*; D011485:Protein Binding; D020449:Repetitive Sequences, Amino Acid
D000911:Antibodies, Monoclonal; D002413:Cations, Divalent; D000939:Epitopes; D006025:Glycosaminoglycans; D008024:Ligands; D009842:Oligopeptides; D011328:Prions; D003300:Copper