Detection and identification of bacterial cell surface proteins by fluorescent labeling

Anaya, Cecilia; Church, Nadia; Lewis, Janina P.

doi:10.1002/pmic.200600175

Cited by 18 publications

(13 citation statements)

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“…The results demonstrate that CyDyes can specifically bind to CSPs, with minimal labeling of cytosolic proteins because CyDyes cannot penetrate the outmost membrane of living cells [43][44][45][46][47]. Moreover, labeling is performed under mild conditions (HBSS, pH 8.5) that are less likely to alter the structure of the algal cell surface [36]. Importantly, cyanine dyes are small and do not alter protein migration in gels [48].…”

Section: Surface Labeling Of a Catenella Dh01 Cellsmentioning

confidence: 91%

“…Moreover, these methods can only address one or a few proteins at one time. Fluorescent labeling of CSPs with Ettan DIGE dyes is a newly developed method and has been widely applied to study the CSPs of various human cell lines [34], bacteria [36] and the monocyte plasma membrane [37]. The results demonstrate that CyDyes can specifically bind to CSPs, with minimal labeling of cytosolic proteins because CyDyes cannot penetrate the outmost membrane of living cells [43][44][45][46][47].…”

Section: Surface Labeling Of a Catenella Dh01 Cellsmentioning

confidence: 95%

“…Recent modifications of traditional DIGE procedures have enabled an elegant examination of CSPs by protein labeling prior to lysis [34,35]. This method has been used to label human cell lines in vitro and in vivo [34], bacterial CSPs [36] and the monocyte plasma membrane [37], and has been shown to be a powerful tool to study CSPs.…”

mentioning

confidence: 99%

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Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling

Wang

Dong

et al. 2012

Chin. Sci. Bull.

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Alexandrium catenella DH01 is a toxic dinoflagellate species that is able to not only produce paralytic shellfish toxins, but also cause harmful algal blooms along the coast of China. In this study, we presented a new protocol for specific labeling and detection of the cell surface proteins (CSPs) of A. catenella DH01 cells using CyDye difference gel electrophoresis (DIGE) fluor minimal dyes. CSPs were identified using two-dimensional gel electrophoresis (2-DE) and MALDI TOF-TOF mass spectrometry (MS). The results showed that the fluorescent cyanine dye Cy3 could specifically label the CSPs of A. catenella DH01, with minimal labeling of intracellular proteins. Among three protein extraction methods evaluated, the Trizol method was the most efficient to extract CSPs with respect to protein spot number and resolution. Forty-one CSPs were separated and identified from A. catenella DH01 by 2-DE, in which 14 were identified in the protein database using MALDI TOF-TOF MS analysis. This work represents the first attempt to investigate the CSPs of A. catenella using the CyDye DIGE fluor dyeing method that provides a potentially important tool for future comprehensive characterization of CSPs and elucidation of the physiological functions of CSPs in dinoflagellates.Alexandrium catenella, cell surface protein, cyanine dye, protein extraction, two-dimensional gel electrophoresis, MALDI TOF-TOF Citation:Li C, Wang D Z, Dong H P, et al. Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling.

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Section: Surface Labeling Of a Catenella Dh01 Cellsmentioning

confidence: 91%

Section: Surface Labeling Of a Catenella Dh01 Cellsmentioning

confidence: 95%

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Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling

Wang

Dong

et al. 2012

Chin. Sci. Bull.

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“…The reaction mixtures were filtered on hot to remove selenium metal and the filtrate was cooled and precipitated by ice water mixture. The precipitated products were filtered, dried, collected, and crystallized from ethanol to give the 4-carbaldehyde compound (5). The data are given in Table (2).…”

Section: Imidazole] (5)mentioning

confidence: 99%

“…Cyanine dyes [1][2][3][4][5][6][7][8][9][10][11][12][13][14] are a class of organic heterocyclic dyes, which have long captured the interest of the scientific community. This is because the class of cyanine dyes has proved to be particularly useful in a diverse and a broad area of science, technology and engineering.…”

Section: Introductionmentioning

confidence: 99%

Synthesis and Photosensitization Evaluation of Some Novel Polyheterocyclic Cyanine Dyes

Shindy¹,

Khalafalla²,

Goma³

et al. 2015

European Reviews of Chemical Research

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New polyheterocyclic compound, namely 4-methyl-6-oxo-2-phenyl-furo [(3,2-d)pyrazole; (  3 ,  2 -d) imidazole] was prepared and oriented to synthesis of some novel monomethine cyanine dyes, dimethine cyanine dyes and mono/di-mixed methine cyanine dyes. The electronic visible absorption spectra of all the synthesized cyanine dyes were investigated in 95% ethanol solution to evaluate their photosensitization properties. Structural identification was carried out via the elemental analysis, visible spectra, mass spectrometry, IR and 1 H NMR spectroscopic data.Keywords: cyanine dyes, synthesis, photosensitization, monomethine cyanine dyes, dimethine cyanine dyes, mixed methine cyanine dyes. IntroductionCyanine dyes [1][2][3][4][5][6][7][8][9][10][11][12][13][14] are a class of organic heterocyclic dyes, which have long captured the interest of the scientific community. This is because the class of cyanine dyes has proved to be particularly useful in a diverse and a broad area of science, technology and engineering. Their uses and applications includes but not limited to color and non color (black and white) photography, in high energy laser and digital image storage, analytical reagents over a wide range of pH of media, indicators for solvent polarity, in biological and biomedical use as molecular probes and as fluorescent dyes commonly used for DNA visualization assays. In addition, cyanine dyes [15][16][17][18][19][20][21][22][23][24][25][26][27][28] are widely used as optical recording materials, thermal writing display, laser printers, laser filters, absorptivity and antimicrobial efficiency, for probing lipophilic environments, halophobic pockets of enzymes and components of supramolecular structures.Taking into account and consideration the above significant benefits of cyanine dyes, in this research paper we prepared new photosensitizers cyanine dyes as new synthesis contribution and spectroscopic investigation in this field and may be used and/or applied in any of the wide mentioned application field of cyanine dyes, particularly as photographic sensitizers in photosensitive material industry, as probes for determining solvent polarity in solution chemistry, and/or as indicators in operations of acid/base titrations.

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Green fluorescent carbon dots as targeting probes for LED‐dependent bacterial killing

et al. 2021

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The emergence of antimicrobial resistance (AMR) represents a significant health and economic challenge worldwide. The slow pace of antibacterial discovery necessitates strategies for optimal use of existing agents, including effective diagnostics able to drive informed prescribing; and development of alternative therapeutic strategies that go beyond traditional small-molecule approaches. Thus, the development of novel probes able to target bacteria for detection and killing, and that can pave the way to effective theranostic strategies, is of great importance.Here we demonstrate that metal-free green-emitting fluorescent carbon dots (FCDs) synthesized from glucosamine⋅HCl and m-phenylenediamine, and featuring 2,5-deoxyfructosazine on a robust amorphous core, can label both Grampositive (Staphylococcus aureus) and Gram-negative (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa) bacterial pathogens within 10 minutes of exposure. Moreover, effective killing of Gram-positive and -negative bacteria can be induced by combining FCD treatment with irradiation by LED light in the visible range. Cell-based, electron microscopy and tandem mass tag (TMT) proteomic experiments indicate that FCD administration in combination with LED exposure gives rise to local heating, ROS production, and membrane-and DNA-damage, suggesting multiple routes to FCD-mediated bacterial killing. Our data identify FCDs as materials that combine facile synthesis from low-cost precursors with labeling and light-dependent killing of clinically important bacterial species, and that thus warrant further exploration as the potential bases for novel theranostics.

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Detection and identification of bacterial cell surface proteins by fluorescent labeling

Cited by 18 publications

References 19 publications

Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling

Proteomics of a toxic dinoflagellate Alexandrium catenella DH01: Detection and identification of cell surface proteins using fluorescent labeling

Synthesis and Photosensitization Evaluation of Some Novel Polyheterocyclic Cyanine Dyes

Green fluorescent carbon dots as targeting probes for LED‐dependent bacterial killing

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