Adsorption of Biological Molecules to a Solid Support for Scanning Probe Microscopy

Müller, Daniel J.; Amrein, M.; Engel, Andréas

doi:10.1006/jsbi.1997.3875

Cited by 286 publications

(210 citation statements)

References 92 publications

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“…The liposomes were stored at −80°C in buffer solution (150 mM NaCl, 20 mM Hepes, pH 7.25). After incubation with 1 μM mM GSDMD and 0.2 μM caspase‐1 for 90 min at 37°C in buffer solution (50 mM NaCl, 100 mM Hepes, 5 mM TCEP, pH 7.4), the liposomes were adsorbed onto freshly cleaved mica in buffer solution (50 mM NaCl, 20 mM Hepes, pH 7.4) (Muller et al , 1997). After an adsorption time of 30 min, the sample was washed several times with the AFM imaging buffer (150 mM NaCl, 20 mM Hepes, pH 7.8) to remove weakly adsorbed protein.…”

Section: Methodsmentioning

confidence: 99%

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

et al. 2016

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Pyroptosis is a lytic type of cell death that is initiated by inflammatory caspases. These caspases are activated within multi‐protein inflammasome complexes that assemble in response to pathogens and endogenous danger signals. Pyroptotic cell death has been proposed to proceed via the formation of a plasma membrane pore, but the underlying molecular mechanism has remained unclear. Recently, gasdermin D (GSDMD), a member of the ill‐characterized gasdermin protein family, was identified as a caspase substrate and an essential mediator of pyroptosis. GSDMD is thus a candidate for pyroptotic pore formation. Here, we characterize GSDMD function in live cells and in vitro. We show that the N‐terminal fragment of caspase‐1‐cleaved GSDMD rapidly targets the membrane fraction of macrophages and that it induces the formation of a plasma membrane pore. In vitro, the N‐terminal fragment of caspase‐1‐cleaved recombinant GSDMD tightly binds liposomes and forms large permeability pores. Visualization of liposome‐inserted GSDMD at nanometer resolution by cryo‐electron and atomic force microscopy shows circular pores with variable ring diameters around 20 nm. Overall, these data demonstrate that GSDMD is the direct and final executor of pyroptotic cell death.

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Section: Methodsmentioning

confidence: 99%

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

et al. 2016

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“…Images occasionally exhibit a loop structure connecting two of the five domains that might correspond to the joining loop 9 . Higher-resolution AFM images have been reported using commercial tips 10 , 11 , but only of densely packed arrays of proteins. When individual molecules have been imaged with these same tips, the resolution is lower 11 .…”

mentioning

confidence: 99%

“…Higher-resolution AFM images have been reported using commercial tips 10 , 11 , but only of densely packed arrays of proteins. When individual molecules have been imaged with these same tips, the resolution is lower 11 .…”

mentioning

confidence: 99%

Growth of nanotubes for probe microscopy tips

Hafner

Cheung

Lieber

1999

Nature

383

228

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“…The ionic strength of the solution will also be much higher close to the surface, depending upon the specific charge of the surface. For unmodified mica in pH 7 solution, this charge is~0.003 C/m 2 (20), corresponding to~1 M ion concentration at the surface (5). Hydration effects are entropy driven, so the effect of the relative hydrophobicity of the surface environment will be enhanced at lower temperature, increasing the driving force for a B to A transition (21).…”

Section: Resultsmentioning

confidence: 99%

Conformational transition in DNA on a cold surface

Feng

Bash

Balagurumoorthy

et al. 2000

Nucleic Acids Research

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The contour length of DNA fragments, deposited and imaged on mica under buffer, was measured as a function of deposition temperature. Extended DNA molecules (on Ni-and silane-treated surfaces) contract rapidly with falling temperature, approaching the contour length of A-DNA at 2°°°°C. The contraction is not unique to a specific sequence and does not occur in solution at 2°°°°C or on a surface at 25°°°°C, indicating that it arises from a combination of low temperature and surface contact. It is probably a consequence of reduced water activity at a cold surface.

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Adsorption of Biological Molecules to a Solid Support for Scanning Probe Microscopy

Cited by 286 publications

References 92 publications

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death

Growth of nanotubes for probe microscopy tips

Conformational transition in DNA on a cold surface

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