TY - JOUR
T1 - Determination of the molecular assembly of actin and actin-binding proteins using photoluminescence
AU - Park, Byeongho
AU - Oh, Seunghee
AU - Jo, Seunghan
AU - Kang, Donyoung
AU - Lim, Juhwan
AU - Jung, Youngmo
AU - Lee, Hyungsuk
AU - Jun, Seong Chan
N1 - Publisher Copyright:
© 2018
PY - 2018/9/1
Y1 - 2018/9/1
N2 - Actin, the most abundant protein in cells, polymerizes into filaments that play key roles in many cellular dynamics. To understand cell dynamics and functions, it is essential to examine the cytoskeleton structure organized by actin and actin-binding proteins. Here, we pave the way for determining the molecular assembly of the actin cytoskeleton using direct photonic in situ analysis, providing the photoluminescence characteristics of actin as a function of filament length and bundling, without labeling. In experiments for monomeric and filamentous actin reconstituted in vitro, structural forms of actin are identified from the peak positions and intensities of photoluminescence. Actin monomers exhibited small intensity emission peaks at 334 nm, whereas filamentous and bundled actin showed a shifted peak at 323 nm with higher intensity. The peak shift was found to be proportional to the length of the actin filament. With probing structural change of actin in various cells in vivo, our study provides an efficient and precise analytical in situ tool to examine the cytoskeleton structure. It will be beneficial for elucidating the mechanism of various cellular functions such as cell migration, differentiation, cytokinesis and adhesion. Furthermore, our technique can be applied to detect the alterations in the cell cytoskeleton that can occur during many pathological processes.
AB - Actin, the most abundant protein in cells, polymerizes into filaments that play key roles in many cellular dynamics. To understand cell dynamics and functions, it is essential to examine the cytoskeleton structure organized by actin and actin-binding proteins. Here, we pave the way for determining the molecular assembly of the actin cytoskeleton using direct photonic in situ analysis, providing the photoluminescence characteristics of actin as a function of filament length and bundling, without labeling. In experiments for monomeric and filamentous actin reconstituted in vitro, structural forms of actin are identified from the peak positions and intensities of photoluminescence. Actin monomers exhibited small intensity emission peaks at 334 nm, whereas filamentous and bundled actin showed a shifted peak at 323 nm with higher intensity. The peak shift was found to be proportional to the length of the actin filament. With probing structural change of actin in various cells in vivo, our study provides an efficient and precise analytical in situ tool to examine the cytoskeleton structure. It will be beneficial for elucidating the mechanism of various cellular functions such as cell migration, differentiation, cytokinesis and adhesion. Furthermore, our technique can be applied to detect the alterations in the cell cytoskeleton that can occur during many pathological processes.
KW - Absorbance
KW - Actin
KW - Cytoskeleton
KW - Label-free determination
KW - Photoluminescence
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UR - http://www.scopus.com/inward/citedby.url?scp=85047651507&partnerID=8YFLogxK
U2 - 10.1016/j.colsurfb.2018.05.043
DO - 10.1016/j.colsurfb.2018.05.043
M3 - Article
C2 - 29852435
AN - SCOPUS:85047651507
SN - 0927-7765
VL - 169
SP - 462
EP - 469
JO - Colloids and Surfaces B: Biointerfaces
JF - Colloids and Surfaces B: Biointerfaces
ER -