Quantification of the number of EP3 receptors on a living CHO cell surface by the AFM

Kim, Hyonchol; Arakawa, Hideo; Hatae, Noriyuki; Sugimoto, Yukihiko; Matsumoto, Osamu; Osada, Toshiya; Ichikawa, Atsushi; Ikai, Atsushi

doi:10.1016/j.ultramic.2005.12.007

Cited by 40 publications

(29 citation statements)

References 32 publications

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“…The force applied to the protein-modified mica surface was kept below 400 pN. The distance force curves and force parameters were obtained according to the methods previously described (39). For antibody neutralization analysis, the protein-modified mica was treated with anti-TLR2 antibodies (diluted 1:1,000 in PBS) for 1 h followed by five washes with PBS to remove unbound antibodies.…”

Section: Methodsmentioning

confidence: 99%

Essential Calcium-binding Cluster of Leptospira LipL32 Protein for Inflammatory Responses through the Toll-like Receptor 2 Pathway

Hsu

et al. 2013

Journal of Biological Chemistry

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Background: LipL32 induces a renal cell inflammatory response through the TLR2-signaling pathway. Results: Ca 2ϩ -binding LipL32 mutants showed attenuated TLR2-mediated inflammatory responses. Conclusion:The Ca 2ϩ -binding cluster of LipL32 is essential in regulating its interaction with TLR2 for subsequent inflammatory response induction. Significance: This investigation provides significant evidence for crucial roles of the Ca 2ϩ -binding cluster of LipL32 for pathogenesis via association with TLR2.

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Section: Methodsmentioning

confidence: 99%

Essential Calcium-binding Cluster of Leptospira LipL32 Protein for Inflammatory Responses through the Toll-like Receptor 2 Pathway

Hsu

et al. 2013

Journal of Biological Chemistry

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“…However, AFM is emerging as a valuable tool to map the distribution of singleproteins on cell surfaces, thereby unraveling the formation and composition of such protein domains. Prominent examples include the mapping of the distribution of prostaglandin receptors on live CHO cells (Kim et al, 2006), the localization of vascular endothelial cadherin-binding sites (Chtcheglova et al, 2007), and the measurement of the organization and stiffness of neuron membrane domains containing glycosylphosphatidylinositol (GPI)-anchored proteins (Roduit et al, 2008). As discussed below, functional protein clusters were recently found to form and increase in size under stress, thereby activating cell signaling and cell adhesion in yeasts.…”

Section: Imaging Protein Clusteringmentioning

confidence: 99%

Atomic force microscopy – looking at mechanosensors on the cell surface

Heinisch

Lipke

Beaussart

et al. 2012

Journal of Cell Science

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SummaryLiving cells use cell surface proteins, such as mechanosensors, to constantly sense and respond to their environment. However, the way in which these proteins respond to mechanical stimuli and assemble into large complexes remains poorly understood at the molecular level. In the past years, atomic force microscopy (AFM) has revolutionized the way in which biologists analyze cell surface proteins to molecular resolution. In this Commentary, we discuss how the powerful set of advanced AFM techniques (e.g. live-cell imaging and single-molecule manipulation) can be integrated with the modern tools of molecular genetics (i.e. protein design) to study the localization and molecular elasticity of individual mechanosensors on the surface of living cells. Although we emphasize recent studies on cell surface proteins from yeasts, the techniques described are applicable to surface proteins from virtually all organisms, from bacteria to human cells.

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“…Considerable efforts have been made to assemble and pattern GFP on various solid surfaces. Some potential applications include photoelectric devices (Choi et al, 2002;Choi et al, 2006;Oh et al, 2004), biosensors (Mazzola et al, 2006), and high sensitive indicators (Kima et al, 2006;Yamada et al, 2004). However, there are few reports for large and uniform GFP monolayer preparation and characterization.…”

Section: Introductionmentioning

confidence: 99%

Assembling and imaging of his‐tag green fluorescent protein on mica surfaces studied by atomic force microscopy and fluorescence microscopy

Liu

et al. 2008

Microscopy Res & Technique

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The adsorption of his-tag green fluorescent protein (GFPH(6)) onto the mica surfaces has been studied by atomic force microscopy (AFM) and laser confocal fluorescence microscopy. By controlling the adsorption conditions, separated single GFPH(6) and GFPH(6) monolayer can be adsorbed and formed on mica surfaces. In present experiments, based on the AFM measurement, we found that the adsorbed GFPH(6) was bound on the mica surface with its beta-sheets. The formed GFPH(6) monolayer on mica surfaces was flat, uniform, and stable. Some applications of the formed monolayer have been demonstrated. The formed monolayer can be used as a substrate for DNA imaging and AFM mechanical lithography.

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Quantification of the number of EP3 receptors on a living CHO cell surface by the AFM

Cited by 40 publications

References 32 publications

Essential Calcium-binding Cluster of Leptospira LipL32 Protein for Inflammatory Responses through the Toll-like Receptor 2 Pathway

Essential Calcium-binding Cluster of Leptospira LipL32 Protein for Inflammatory Responses through the Toll-like Receptor 2 Pathway

Atomic force microscopy – looking at mechanosensors on the cell surface

Assembling and imaging of his‐tag green fluorescent protein on mica surfaces studied by atomic force microscopy and fluorescence microscopy

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