Phospholipids enhance nucleation but not elongation of apolipoprotein C-II amyloid fibrils.
Amyloid fibrils and their oligomeric intermediates accumulate in several age-related diseases where their presence is considered to play an active role in disease progression. A common characteristic of amyloid fibril formation is an initial lag phase indicative of a nucleation-elongation mechanism for fibril assembly. We have investigated fibril formation by human apolipoprotein (apo) C-II. ApoC-II readily forms amyloid fibrils in a lipid-dependent manner via an initial nucleation step followed by fibril elongation, breaking, and joining. We used fluorescence techniques and stopped-flow analysis to identify the individual kinetic steps involved in the activation of apoC-II fibril formation by the short-chain phospholipid dihexanoyl phosphatidylcholine (DHPC). Submicellar DHPC activates fibril formation by promoting the rapid formation of a tetrameric species followed by a slow isomerisation that precedes monomer addition and fibril growth. Global fitting of the concentration dependence of apoC-II fibril formation showed that DHPC increased the overall tetramerisation constant from 7.5 x 10(-13) to 1.2 x 10(-6) microM(-3) without significantly affecting the rate of fibril elongation, breaking, or joining. Studies on the effect of DHPC on the free pool of apoC-II monomer and on fibril formation by cross-linked apoC-II dimers further demonstrate that DHPC affects nucleation but not elongation. These studies demonstrate the capacity of small lipid compounds to selectively target individual steps in the amyloid fibril forming pathway.
Ryan
Timothy M
TM
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Melbourne, Victoria 3010, Australia.
Teoh
Chai L
CL
Griffin
Michael D W
MD
Bailey
Michael F
MF
Schuck
Peter
P
Howlett
Geoffrey J
GJ
D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism; D053304:Apolipoprotein C-II / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D019281:Dimerization; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D010713:Phosphatidylcholines / Q000737:chemistry*
D000682:Amyloid; D053304:Apolipoprotein C-II; D010713:Phosphatidylcholines; C032166:1,2-hexanoylphosphatidylcholine