Studying molecular interactions in the intact organism: fluorescence correlation spectroscopy in the living zebrafish embryo

Dawes, Michael L; Soeller, Christian; Scholpp, Steffen

doi:10.1007/s00418-020-01930-5

Cited by 12 publications

(8 citation statements)

References 103 publications

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“…Therefore, in order to quantify protein-protein interactions directly at the PM of living cells, we performed fluorescence fluctuation spectroscopy experiments under physiological conditions. Such experimental approaches (i.e., sF(C)CS and (cc)N&B) provide information regarding the oligomeric state, surface concentration, hetero-interactions and dynamics in complex biological systems (51,52,(63)(64)(65)(66).…”

Section: Discussionmentioning

confidence: 99%

Influenza A M2 recruits M1 to the plasma membrane: a fluorescence fluctuation microscopy study

Petrich

Dunsing

Bobone

et al. 2021

Preprint

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Influenza A virus (IAV) is a respiratory pathogen that causes seasonal epidemics and occasional pandemics of severe illnesses with significant mortality. One of the most abundant proteins in IAV particles is the matrix protein 1 (M1), which is essential for the structural stability of the virus. M1 organizes virion assembly and budding at the plasma membrane (PM), where it can interact with other viral components and cellular membrane factors (i.e. lipids and host proteins). Of interest, the recruitment of M1 to the PM as well as its interaction with the other viral envelope proteins (hemagglutinin (HA), neuraminidase, matrix protein 2 (M2)) is controversially discussed in previous studies. Therefore, we used fluorescence fluctuation microscopy techniques (i.e. (cross-correlation) number and brightness, and scanning fluorescence cross-correlation spectroscopy) to quantify the oligomeric state of M1 and its interaction with other viral proteins in co-transfected as well as infected cells. Our results indicate that M1 is recruited to the PM by M2, as a consequence of the strong interaction between the two proteins. In contrast, only a weak interaction between M1 and HA was observed. M1-HA interaction occurred only in the case that M1 was already bound to the PM. We therefore conclude that M2 initiates the assembly of IAV by recruiting M1 to the PM, possibly allowing its further interaction with other viral proteins.

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Section: Discussionmentioning

confidence: 99%

Influenza A M2 recruits M1 to the plasma membrane: a fluorescence fluctuation microscopy study

Petrich

Dunsing

Bobone

et al. 2021

Preprint

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“…In-cell NMR serves as a promising approach to provide structural and dynamics data on protein-protein interaction and protein-ligand interaction systems in cellular environments [ 45 , 49 , 151 ]. Generally, in-cell NMR can be carried out in bacteria, yeast, sf9, frog oocytes, zebrafish embryo, and human cells without further sample purification, and it has become a remarkable tool for drug discovery [ 45 , 152 , 153 , 154 , 155 , 156 , 157 , 158 ]. Multiple cases related to the applications of in-cell NMR in drug discovery have been reported.…”

Section: In-cell Nmrmentioning

confidence: 99%

Applications of Solution NMR in Drug Discovery

Shi

Zhang

2021

Molecules

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During the past decades, solution nuclear magnetic resonance (NMR) spectroscopy has demonstrated itself as a promising tool in drug discovery. Especially, fragment-based drug discovery (FBDD) has benefited a lot from the NMR development. Multiple candidate compounds and FDA-approved drugs derived from FBDD have been developed with the assistance of NMR techniques. NMR has broad applications in different stages of the FBDD process, which includes fragment library construction, hit generation and validation, hit-to-lead optimization and working mechanism elucidation, etc. In this manuscript, we reviewed the current progresses of NMR applications in fragment-based drug discovery, which were illustrated by multiple reported cases. Moreover, the NMR applications in protein-protein interaction (PPI) modulators development and the progress of in-cell NMR for drug discovery were also briefly summarized.

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“…After a fluorescently labeled molecule binds to its interaction partner, its mobility is slowed, which is reflected in its fluctuation rate (Langowski 2008). FCS can also determine the concentration and mobility (diffusion coefficient) of free interacting proteins (Langowski 2008;Dawes et al 2020). FCS can be applied in vivo for qualitative confirmation of PPIs, for example, as described in the interaction of human immunodeficiency virus type 1 integrase with lens epithelium-derived growth factor/transcription co-activator p75 (Maertens et al 2005).…”

Section: Fluorescence Correlation Spectroscopy (Fcs) and Fluorescence Cross-correlation Spectroscopy (Fccs)mentioning

confidence: 99%

Technologies for the identification and validation of protein-protein interactions

Pichlerova¹,

Hanes²

2021

gpb

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Proteins are large molecules that play essential roles in all living organisms. In most molecular processes in each cell, proteins usually do not function alone but through physiological interactions with various ligands. The most common interacting molecules for proteins are other proteins, and they act together by protein-protein interactions (PPIs) to create larger protein complexes. The impairment of physiological PPIs or establishing PPIs with pathological proteins often leads to the development of diseases. To bring insights on the knowledge about the physiological functions of proteins in biological processes, and to understand the development and pathogenesis of diseases, numerous qualitative and quantitative methods have been developed. In this review, we summarize the most commonly used methods for studying PPIs, and discuss their advantages and drawbacks.

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Studying molecular interactions in the intact organism: fluorescence correlation spectroscopy in the living zebrafish embryo

Cited by 12 publications

References 103 publications

Influenza A M2 recruits M1 to the plasma membrane: a fluorescence fluctuation microscopy study

Influenza A M2 recruits M1 to the plasma membrane: a fluorescence fluctuation microscopy study

Applications of Solution NMR in Drug Discovery

Technologies for the identification and validation of protein-protein interactions

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