Surface‐Charge Differentiation of Streptavidin and Avidin by Atomic Force Microscopy–Force Spectroscopy

Almonte, Lisa; López-Elvira, Elena; Baró, A. M.

doi:10.1002/cphc.201402234

Cited by 31 publications

(34 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…These estimates are consistent with the results obtained from STM images. Numerous reports have shown that protein hydration and tip radius artifacts affect solution-based STM and AFM images [34,35,49,50]. Although we successfully resolved sub-molecular features, greater molecule size reflected is a limitation of our method.…”

Section: Discussioncontrasting

confidence: 58%

“…Its crystal structure was determined by multi-wavelength anomalous diffraction [33]. Streptavidin images obtained using scanning probe microscopes appear as round spots without sub-molecular details [34][35][36][37]. Figure 2(a) shows an STM image of streptavidin molecules that resembles the crystal structure, but are not identical.…”

Section: Sub-molecular Features Observed On Streptavidin and Antibodymentioning

confidence: 99%

See 1 more Smart Citation

Sub-molecular features of single proteins in solution resolved with scanning tunneling microscopy

Wang

Zhang

et al. 2016

Nano Res.

View full text Add to dashboard Cite

These authors contributed equally to this work. ABSTRACTScanning tunneling microscopy (STM) can be used to image individual biological molecules, such as proteins, in vacuum or air. This requires sample dehydration and thus may not reflect the native state of the molecule. Extensive efforts have been made to image single proteins in solution using STM; however, the images have revealed only round or oval shapes with no sub-molecular details. Here, we present the sub-molecular features of streptavidin proteins under physiological conditions using a homebuilt low-leakage-current and highstability liquid phase STM. The N-lobe, C-lobe, and C-terminal tail of the epidermal growth factor receptor kinase domains were also resolved in solution. Our results demonstrate that the structure, morphology, and dynamics of a protein molecule can be examined under physiological conditions by the STM.

show abstract

Section: Discussioncontrasting

confidence: 58%

Section: Sub-molecular Features Observed On Streptavidin and Antibodymentioning

confidence: 99%

Sub-molecular features of single proteins in solution resolved with scanning tunneling microscopy

Wang

Zhang

et al. 2016

Nano Res.

View full text Add to dashboard Cite

show abstract

“…The pI of surfacebound streptavidin has been measured using Surface Force Apparatus (SFA) and Atomic Force Microscopy which demonstrated a pI of 5.0 ± 0.5. 79,83 Experimental determination of streptavidin pI is discussed further in ESI section 4. † Other biomolecules.…”

Section: Streptavidin Biochemistrymentioning

confidence: 99%

Field-effect sensors – from pH sensing to biosensing: sensitivity enhancement using streptavidin–biotin as a model system

Lowe

Sun

Zeimpekis

et al. 2017

Analyst

128

107

View full text Add to dashboard Cite

a Field-Effect Transistor sensors (FET-sensors) have been receiving increasing attention for biomolecular sensing over the last two decades due to their potential for ultra-high sensitivity sensing, label-free operation, cost reduction and miniaturisation. Whilst the commercial application of FET-sensors in pH sensing has been realised, their commercial application in biomolecular sensing (termed BioFETs) is hindered by poor understanding of how to optimise device design for highly reproducible operation and high sensitivity. In part, these problems stem from the highly interdisciplinary nature of the problems encountered in this field, in which knowledge of biomolecular-binding kinetics, surface chemistry, electrical double layer physics and electrical engineering is required. In this work, a quantitative analysis and critical review has been performed comparing literature FET-sensor data for pH-sensing with data for sensing of biomolecular streptavidin binding to surface-bound biotin systems. The aim is to provide the first systematic, quantitative comparison of BioFET results for a single biomolecular analyte, specifically streptavidin, which is the most commonly used model protein in biosensing experiments, and often used as an initial proofof-concept for new biosensor designs. This novel quantitative and comparative analysis of the surface potential behaviour of a range of devices demonstrated a strong contrast between the trends observed in pH-sensing and those in biomolecule-sensing. Potential explanations are discussed in detail and surfacechemistry optimisation is shown to be a vital component in sensitivity-enhancement. Factors which can influence the response, yet which have not always been fully appreciated, are explored and practical suggestions are provided on how to improve experimental design.

show abstract

“…Additionally, using these techniques it is difficult to assess the properties of functional viral particles in physiological conditions. The density of charge of other biomolecules, such as single dsDNA molecules 13 , supported lipid bilayers 14 , and the avidin-streptavidin system 15 have also been measured. The subsequent refinement of this technique allowed the measurement of the electrical charge of biomolecules forming monolayers, such as purple membrane 12 resulting in a density of charge of -0.312 e 0 /nm 2 , where e 0 is the absolute value of the electron charge (1 C/m 2 = 6.24e 0 /nm 2 ).…”

Section: Introductionmentioning

confidence: 99%

Quantitative nanoscale electrostatics of viruses

et al. 2015

View full text Add to dashboard Cite

Electrostatics is one of the fundamental driving forces of the interaction between biomolecules in solution. In particular, the recognition events between viruses and host cells are dominated by both specific and non-specific interactions and the electric charge of viral particles determines the electrostatic force component of the latter. Here we probe the charge of individual viruses in liquid milieu by measuring the electrostatic force between a viral particle and the Atomic Force Microscope tip. The force spectroscopy data of co-adsorbed ϕ29 bacteriophage proheads and mature virions, adenovirus and minute virus of mice capsids is utilized for obtaining the corresponding density of charge for each virus. The systematic differences of the density of charge between the viral particles are consistent with the theoretical predictions obtained from X-ray structural data. Our results show that the density of charge is a distinguishing characteristic of each virus, depending crucially on the nature of the viral capsid and the presence/absence of the genetic material.

show abstract

Surface‐Charge Differentiation of Streptavidin and Avidin by Atomic Force Microscopy–Force Spectroscopy

Cited by 31 publications

References 60 publications

Sub-molecular features of single proteins in solution resolved with scanning tunneling microscopy

Sub-molecular features of single proteins in solution resolved with scanning tunneling microscopy

Field-effect sensors – from pH sensing to biosensing: sensitivity enhancement using streptavidin–biotin as a model system

Quantitative nanoscale electrostatics of viruses

Contact Info

Product

Resources

About