Abstract:
:Marvelous background rejection in total internal reflection fluorescence microscopy (TIR-FM) has made it possible to visualize single-fluorophores in living cells. Cell signaling proteins including peptide hormones, membrane receptors, small G proteins, cytoplasmic kinases as well as small signaling compounds have been conjugated with single chemical fluorophore or tagged with green fluorescent proteins and visualized in living cells. In this review, the reasons why single-molecule analysis is essential for studies of intracellular protein systems such as cell signaling system are discussed, the instrumentation of TIR-FM for single-molecule imaging in living cells is explained, and how single molecule visualization has been used in cell biology is illustrated by way of two examples: signaling of epidermal growth factor in mammalian cells and chemotaxis of Dictyostelium amoeba along a cAMP gradient. Single-molecule analysis is an ideal method to quantify the parameters of reaction dynamics and kinetics of unitary processes within intracellular protein systems. Knowledge of these parameters is crucial for the understanding of the molecular mechanisms underlying intracellular events, thus single-molecule imaging in living cells will be one of the major technologies in cellular nanobiology.
journal_name
Cell Struct Functjournal_title
Cell structure and functionauthors
Sako Y,Uyemura Tdoi
10.1247/csf.27.357keywords:
subject
Has Abstractpub_date
2002-10-01 00:00:00pages
357-65issue
5eissn
0386-7196issn
1347-3700journal_volume
27pub_type
杂志文章,评审abstract::The unique Physarum lysophosphatidic acid, PHYLPA, having a cyclopropane in the fatty acid moiety and a cyclic phosphate at C-2 and C-3 positions of the glycerol, inhibited proliferation of human fibroblast cells, TIG-3 and TIG-7, which were cultured in a chemically defined (serum-free) medium. The cells at S- and M-p...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.18.363
更新日期:1993-10-01 00:00:00
abstract::Synapsin I, one of the major synaptic proteins, is thought to associate with synaptic vesicles and to play a regulatory role in neurotransmitter release. In mature neurons, it is concentrated almost exclusively in presynaptic nerve endings. Here, we studied the subcellular localization of synapsin I during the develop...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.15.329
更新日期:1990-12-01 00:00:00
abstract::Two-dimensional gel electrophoresis revealed two protein spots, a (molecular weight 70,000, pI 4.6) and b (molecular weight 69,000, pI 4.4) in PC-12 cells (rat pheochromocytoma cells). When the cells were induced to differentiate with nerve growth factor, the amount of protein in spot a, and later spot b increased wit...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.127
更新日期:1987-02-01 00:00:00
abstract::Epithelial tubules, consisting of the epithelial cell sheet with a central lumen, are the basic structure of many organs. Mechanical forces play an important role in epithelial tubulogenesis; however, little is known about the mechanisms controlling the mechanical forces during epithelial tubule morphogenesis. Solo (a...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.18010
更新日期:2018-06-02 00:00:00
abstract::Cholera toxin binding sites in frozen sections of rat thymus were studied by indirect immunofluorescence and were found to be localized selectively in the medulla of thymic lobules. By flow cytometric analysis, two populations of thymocytes differing in size and reactivity towards cholera toxin were detected; the smal...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.339
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abstract::A large number of Rab/Ypt-family small GTPases have been identified from higher plants. While some of them can complement yeast ypt mutants, the expression of Arabidopsis Ara4 protein aggravated the growth defect of a subset of ypt mutants, probably because of the titration of common regulator(s) of yeast Ypt proteins...
journal_title:Cell structure and function
pub_type: 杂志文章
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abstract::Two types of DNA ligase, I and II, have been purified approximately 4,000-fold from mouse testes and 500-fold from nuclei of mouse spermatocytes. DNA ligase I and II consisted of single polypeptides with molecular weights of 95,000 and 65,000, respectively, according to the estimation by SDS-polyacrylamide gel electro...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.15.67
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abstract::The in vivo structure of the smooth endoplasmic reticulum (ER) was visualized in rat and mouse cerebellar Purkinje cells by using quick-freezing techniques followed by freeze-substitution for ultrathin-sectioning or freeze-fracturing and deep-etching for replicas. High magnification electron microscopy of the ultrathi...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.23.373
更新日期:1998-12-01 00:00:00
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journal_title:Cell structure and function
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更新日期:2016-07-30 00:00:00
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journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.22.545
更新日期:1997-10-01 00:00:00
abstract::Human keratinocytes have been serially cultivated in low (0.015 mM) and high (1.8 mM) calcium containing medium. The calcium concentration of the growth medium significantly influenced the cell growth period in vitro. Cells grown in low calcium medium underwent 35-40 population doublings over 16-17 passages, while cel...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.539
更新日期:1987-12-01 00:00:00
abstract::Microtubules are intrinsically dynamic polymers. Two kinds of dynamic behaviors, dynamic instability and treadmilling, are important for microtubule function in cells. Both dynamic behaviors appear to be tightly regulated, but the cellular molecules and the mechanisms responsible for the regulation remain largely unex...
journal_title:Cell structure and function
pub_type: 杂志文章,评审
doi:10.1247/csf.24.329
更新日期:1999-10-01 00:00:00
abstract::Hepatic stellate cells (HSCs) were changed in their morphology, proliferative activity, and functions by culturing on type I collagen gel, as compared to the culture on polystyrene surface. HSCs have been found to produce extracellular matrix components and matrix metalloproteinases (MMPs). In this study, we have asse...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.24.255
更新日期:1999-10-01 00:00:00
abstract::G0-arrested human diploid fibroblasts, TIG-1, was stimulated to induce DNA synthesis by serum, epidermal growth factor (EGF), colchicine, colcemid, or 12-O-tetradecanoylphorbol-13-acetate (TPA). The induction of DNA synthesis was mediated by protein kinase C (PKC) when stimulated with TPA but not when stimulated with ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.16.73
更新日期:1991-02-01 00:00:00
abstract::Genomics and proteomics studies in Chlamydomonas have revealed that an axoneme is composed of 200-600 types of proteins, including uncharacterized proteins collectively named flagellar-associated proteins (FAPs). Nine FAPs contain the EF-hand motif; however, they have not yet been well characterized. To find component...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.17023
更新日期:2018-02-16 00:00:00
abstract::DNA-dependent protein kinase (DNA-PK), consisting of the 470-kDa catalytic component (DNA-PKcs) and the DNA-binding regulatory component Ku protein (p70/p80), catalyzes phosphorylation of a variety of DNA replication/transcription/repair factors in the presence of double-stranded DNA. In the resting states of human pe...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.22.585
更新日期:1997-12-01 00:00:00
abstract::To construct recombinant retroviruses with only a single active promoter, we introduced point mutations into the TATA box region of the 3'-LTR, and successfully obtained high-titer virus with sufficient self-inactivating activity. However, the viral titer could not be determined by the number of G418 resistant colonie...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.177
更新日期:1995-04-01 00:00:00
abstract::The relative fibrin-binding, fibrinolytic and fibrinogenolytic properties of single-chain pro-urokinase, an inactive proenzyme form of human urokinase purified from cultured human kidney cells, and urokinase were compared. The affinity of single-chain pro-urokinase for fibrin was much higher than that of urokinase. In...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.10.151
更新日期:1985-06-01 00:00:00
abstract::The involvement of actin filaments from the host cell on the process of invasion of trypomastigote forms of Trypanosma cruzi was analyzed in seven different cell lines. Prior incubation of all cell lines with cytochalasin D, under conditions which interfere with actin filaments, markedly inhibited parasite internaliza...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.27.91
更新日期:2002-04-01 00:00:00
abstract::Cells of Euglena gracilis Klebs var. bacillaris Cori mutant W3BUL grown in darkness on Hutner's pH 3.5 medium without agitation accumulate wax ester. These cells have undeveloped proplastid remnants characteristic of this mutant. If these cells are transferred to an inorganic medium and bubbled with 2-3% CO2 in air, t...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.12.453
更新日期:1987-10-01 00:00:00
abstract::In this study, we measured hydrogen peroxide (H2O2) release as one of the functions of mature eosinophils, and utilized it as a quantitative index. We demonstrated that 1) the human eosinophilic leukemia cell line, EoL-1, did not release H2O2 when stimulated with phorbol myristate acetate (PMA), but after culturing wi...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.18.125
更新日期:1993-04-01 00:00:00
abstract::Here we report a new method of isolating epidermal desmosomes from Xenopus laevis, and a major constituent of desmosomes designated as Xenopus desmogleins (XDsg). Isolation of desmosomes from Xenopus laevis epidermis was carried out by a two step-incubation with different concentrations of NP-40. After discontinuous s...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.29.17
更新日期:2004-02-01 00:00:00
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journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.14.173
更新日期:1989-04-01 00:00:00
abstract::Here we describe the design and synthesis of a bifunctional two-photon fluorescence probe, N,N'-dimethyl-4,4'-(biphenyl-2,1-ethenediyl)dipyridinium hexafluorophosphate (BP6). HeLa, Hek293, and Paramecium caudatum cells were stained with BP6. BP6 accumulated on the mitochondria of all three cell types when the mitocho...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.14006
更新日期:2014-01-01 00:00:00
abstract::The morphology and intramembranous particle (IMP) distribution of spermatozoa were investigated in four species of cyprinid fishes, Gnathopogon elongatus elongatus, Pseudogobio esocinus, Acheilognathus lanceolatus and Puntius tetrazona. In particular, observation of IMP distribution was conducted as an aspect of compa...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.293
更新日期:1995-08-01 00:00:00
abstract::A monoclonal type M-immunoglobulin (IgM) was generated in mice against a nuclease-urea extract of HeLa metaphase chromosomes. This antibody stains metaphase chromosomes from a variety of mammalian cultured cell types by indirect immunofluorescence. Antibody 12C7 reacts by western transfer technique with histone H1 in ...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.16.323
更新日期:1991-08-01 00:00:00
abstract::Staurosporine has been reported to cause arrest of cells in G1 phase at low concentration and in G2 phase at high concentration. This raises the question of why the effects of staurosporine on the cell cycle depend on the applied concentration. In order to verify these multiple functions of staurosporine in Meth-A cel...
journal_title:Cell structure and function
pub_type: 杂志文章
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更新日期:1999-12-01 00:00:00
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journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.20.345
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abstract::Caffeine has been shown to enhance the lethal effect of DNA-damaging agents in mammalian cells, and the potentiation by caffeine of this effect is generally interpreted as the result of inhibition by caffeine of the repair of damaged DNA. However, the mechanism by which caffeine enhances the lethal effect of DNA-damag...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.10.405
更新日期:1985-12-01 00:00:00
abstract::Autophagy is a starvation response in eukaryotes by which the cell delivers cytoplasmic components to the vacuole for degradation, and is mediated by a double membrane structure called the autophagosome. We have previously proposed that the specific combination of COPII like components, including Sec24p, is required f...
journal_title:Cell structure and function
pub_type: 杂志文章
doi:10.1247/csf.28.49
更新日期:2003-02-01 00:00:00