TY - JOUR
T1 - In situ monitored vortex fluidic-mediated protein refolding/unfolding using an aggregation-induced emission bioprobe
AU - Hu, Qi
AU - Hu, Haozhen
AU - Zhang, Xinyi
AU - Fan, Kyle
AU - Hong, Yuning
AU - Raston, Colin L.
AU - Tang, Youhong
PY - 2021/7/2
Y1 - 2021/7/2
N2 - Protein folding is important for protein homeostasis/proteostasis in the human body. We have established the ability to manipulate protein unfolding/refolding for β-lactoglobulin using the induced mechanical energy in the thin film microfluidic vortex fluidic device (VFD) with monitoring as such using an aggregation-induced emission luminogen (AIEgen), TPE-MI. When denaturant (guanidine hydrochloride) is present with β-lactoglobulin, the VFD accelerates the denaturation reaction in a controlled way. Conversely, rapid renaturation of the unfolded protein occurs in the VFD in the absence of the denaturant. The novel TPE-MI reacts with exposed cysteine thiol when the protein unfolds, as established with an increase in fluorescence intensity. TPE-MI provides an easy and accurate way to monitor the protein folding, with comparable results established using conventional circular dichroism. The controlled VFD-mediated protein folding coupled with in situ bioprobe AIEgen monitoring is a viable methodology for studying the denaturing of proteins.
AB - Protein folding is important for protein homeostasis/proteostasis in the human body. We have established the ability to manipulate protein unfolding/refolding for β-lactoglobulin using the induced mechanical energy in the thin film microfluidic vortex fluidic device (VFD) with monitoring as such using an aggregation-induced emission luminogen (AIEgen), TPE-MI. When denaturant (guanidine hydrochloride) is present with β-lactoglobulin, the VFD accelerates the denaturation reaction in a controlled way. Conversely, rapid renaturation of the unfolded protein occurs in the VFD in the absence of the denaturant. The novel TPE-MI reacts with exposed cysteine thiol when the protein unfolds, as established with an increase in fluorescence intensity. TPE-MI provides an easy and accurate way to monitor the protein folding, with comparable results established using conventional circular dichroism. The controlled VFD-mediated protein folding coupled with in situ bioprobe AIEgen monitoring is a viable methodology for studying the denaturing of proteins.
KW - Aggregation induced emission
KW - Protein folding/unfolding
KW - Vortex fluidic device
UR - http://www.scopus.com/inward/record.url?scp=85111095499&partnerID=8YFLogxK
UR - http://purl.org/au-research/grants/ARC/DP200101106
U2 - 10.3390/molecules26144273
DO - 10.3390/molecules26144273
M3 - Article
AN - SCOPUS:85111095499
SN - 1420-3049
VL - 26
JO - Molecules
JF - Molecules
IS - 14
M1 - 4273
ER -