Dimerization of beta 2-adrenergic receptor (β-AR) has been observed across various physiologies. However, the function of dimeric β-AR is still elusive. Here, we revealed that dimerization of β-AR is responsible for the constitutive activity of β-AR generating inverse agonism.
View Article and Find Full Text PDFMulticolor fluorescence imaging is a powerful tool visualizing the spatiotemporal relationship among biomolecules. Here, we report that commonly employed organic dyes exhibit a blue-conversion phenomenon, which can produce severe multicolor image artifacts leading to false-positive colocalization by invading predefined spectral windows, as demonstrated in the case study using EGFR and Tensin2. These multicolor image artifacts become much critical in localization-based superresolution microscopy as the blue-converted dyes are photoactivatable.
View Article and Find Full Text PDFSingle-molecule localization microscopy (SMLM) has allowed the observation of various molecular structures in cells beyond the diffraction limit using organic dyes. In principle, the SMLM resolution depends on the precision of photoswitching fluorophore localization, which is inversely correlated with the square root of the number of photons released from the individual fluorophores. Thus, increasing the photon number by using highly bright fluorophores, such as quantum dots (QDs), can theoretically fundamentally overcome the current resolution limit of SMLM.
View Article and Find Full Text PDFVarious repertoires of membrane protein interactions determine cellular responses to diverse environments around cells dynamically in space and time. Current assays, however, have limitations in unraveling these interactions in the physiological states in a living cell due to the lack of capability to probe the transient nature of these interactions on the crowded membrane. Here, we present a simple and robust assay that enables the investigation of transient protein interactions in living cells by using the single-molecule diffusional mobility shift assay (smDIMSA).
View Article and Find Full Text PDFInteractions between membrane proteins are poorly understood despite their importance in cell signaling and drug development. Here, we present a co-immunoimmobilization assay (Co-II) enabling the direct observation of membrane protein interactions in single living cells that overcomes the limitations of currently prevalent proximity-based indirect methods. Using Co-II, we investigated the transient homodimerizations of epidermal growth factor receptor (EGFR) and beta-2 adrenergic receptor (β2-AR) in living cells, revealing the differential regulation of these receptors' dimerizations by molecular conformations and microenvironment in a plasma membrane.
View Article and Find Full Text PDFCellular processes occur through the orchestration of multi-step molecular reactions. Reaction progress kinetic analysis (RPKA) can provide the mechanistic details to elucidate the multi-step molecular reactions. However, current tools have limited ability to simultaneously monitor dynamic variations in multiple complex states at the single molecule level to apply RPKA in living cells.
View Article and Find Full Text PDFWith a purpose to find out natural transition of endemicity of Malayan filariasis in inland Korea, a survey was conducted in June 1980 in Isan-Myeon of Yongpung-Gun (former Yongju-Gun) where an epidemiological investigation had been carried out in 1973 without any control activities such as chemotherapy. Five sample villages were surveyed for microfilaremia by 20 microliter night blood examination among inhabitants and the results of the surveys conducted in 1973 and 1980 were compared to determine natural transition of the endemicity of malayan filariasis during the period of the last 7 years. 1.
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