Amyloid Formation in Nanoliter Droplets.
amyloid formation
atomic force microscopy
fluorescence assay
lysozyme
nanoliter droplet
polymorphism
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
13 May 2022
13 May 2022
Historique:
received:
30
03
2022
revised:
11
05
2022
accepted:
12
05
2022
entrez:
28
5
2022
pubmed:
29
5
2022
medline:
1
6
2022
Statut:
epublish
Résumé
Processes that monitor the nucleation of amyloids and characterize the formation of amyloid fibrils are vital to medicine and pharmacology. In this study, we observe the nucleation and formation of lysozyme amyloid fibrils using a facile microfluidic system to generate nanoliter droplets that can control the flow rate and movement of monomer-in-oil emulsion droplets in a T-junction microchannel. Using a fluorescence assay, we monitor the nucleation and growth process of amyloids based on the volume of droplets. Using the microfluidic system, we demonstrate that the lag phase, which is vital to amyloid nucleation and growth, is reduced at a lower droplet volume. Furthermore, we report a peculiar phenomenon of high amyloid formation at the edge of a bullet-shaped droplet, which is likely due to the high local monomer concentration. Moreover, we discovered that amyloid fibrils synthesized in the nanoliter droplets are shorter and thicker than fibrils synthesized from a bulk solution via the conventional heating method. Herein, a facile procedure to observe and characterize the nucleation and growth of amyloid fibrils using nanoliter droplets is presented, which is beneficial for investigating new features of amyloid fibril formation as an unconventional synthetic method for amyloid fibrils.
Identifiants
pubmed: 35628295
pii: ijms23105480
doi: 10.3390/ijms23105480
pmc: PMC9143811
pii:
doi:
Substances chimiques
Amyloid
0
Amyloidogenic Proteins
0
Emulsions
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : National Research Foundation of Korea
ID : NRF-2020R1A2C2102262
Organisme : National Research Foundation of Korea
ID : KMDF_PR_20200901_0127-01
Organisme : National Research Foundation of Korea
ID : 2021R1A4A1028969
Organisme : Korea University
ID : BK21 FOUR
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