Detection of Conformational Changes in an Immobilized Protein Using Surface Plasmon Resonance

Sota, Hiroyuki; Hasegawa, Yukio; Iwakura, Masahiro

doi:10.1021/ac9713666

Cited by 170 publications

(123 citation statements)

References 29 publications

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“…They observed that the SPR signal of the tethered protein and the molar ellipticity of dihydrofolate reductase in solution responded similarly to pH changes. Combined with tethered protein sensor surface measurements in the pH range of 0.12-7.80, they postulated that their observations were "consistent with the interpretation that changes in the SPR signal reflect conformational changes occurring during acid denaturation" (25).…”

Section: Introductionmentioning

confidence: 75%

“…Although we, and others, have postulated the origin of the free-solution signal, no explicit explanation for the physical phenomenon has emerged. Here, we capitalize upon the pioneering observations by Sota and others including Pitner and Koch (25)(26)(27)(28), using techniques typically thought to be insensitive to bulk RI changes (25) that couple energy into an immobilized sensor surface layer, which suggest that a theory based purely on dη/dC considerations does not adequately describe the response for optical methods performed in free solution. The background necessary to support this supposition is provided below and in SI Appendix.…”

Section: Introductionmentioning

confidence: 99%

“…The model described above and in SI Appendix used to define the response for RI (bio)sensing methods, does not take into account the possibility that the signal may be impacted by conformation and hydration changes upon chemical or biochemical transformation (binding, folding, shedding, or gaining waters of hydration). To our knowledge, Sota et al (25) were the first to question this supposition by reporting the detection of conformational changes in an immobilized protein using an SPR biosensor. They observed that the SPR signal of the tethered protein and the molar ellipticity of dihydrofolate reductase in solution responded similarly to pH changes.…”

Section: Introductionmentioning

confidence: 99%

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Origin and prediction of free-solution interaction studies performed label-free

Bornhop

Kammer

Kussrow

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Interaction/reaction assays have led to significant scientific discoveries in the biochemical, medical, and chemical disciplines. Several fundamental driving forces form the basis of intermolecular and intramolecular interactions in chemical and biochemical systems (London dispersion, hydrogen bonding, hydrophobic, and electrostatic), and in the past three decades the sophistication and power of techniques to interrogate these processes has developed at an unprecedented rate. In particular, label-free methods have flourished, such as NMR, mass spectrometry (MS), surface plasmon resonance (SPR), biolayer interferometry (BLI), and backscattering interferometry (BSI), which can facilitate assays without altering the participating components. The shortcoming of most refractive index (RI)-based label-free methods such as BLI and SPR is the requirement to tether one of the interaction entities to a sensor surface. This is not the case for BSI. Here, our hypothesis is that the signal origin for freesolution, label-free determinations can be attributed to conformation and hydration-induced changes in the solution RI. We propose a model for the free-solution response function (FreeSRF) and show that, when quality bound and unbound structural data are available, FreeSRF correlates well with the experiment (R 2 > 0.99, Spearman rank correlation coefficients >0.9) and the model is predictive within ∼15% of the experimental binding signal. It is also demonstrated that a simple mass-weighted dη/dC response function is the incorrect equation to determine that the change in RI is produced by binding or folding event in free solution.backscattering interferometry | assay methodology | molecular interactions | conformation change | hydration change C ontemporary assays enabling single-molecule detection (1,2) have accelerated the sequencing of the human genome (3) and facilitated imaging with extraordinary resolution without labels (4). To most closely approximate the natural state, an interaction assay methodology would interrogate the processes (reaction, molecular interaction, protein folding event, etc.) without perturbation. Label-free chemical and biochemical investigations (5, 6) transduce the desired signal without an exogenous label (fluorescent, radioactive, or otherwise) representing an essential step toward this goal. Many label-free methods require one of the interacting species to be either tethered or immobilized to the sensor surface, introducing a potential perturbation to the natural state of the species (7,8). However, back-scattering interferometry (BSI) is a free-solution label-free technique with the added benefit of sensitivity that rivals fluorescence (9). There are other techniques performed in free solution, such as MS (10, 11) and NMR (12,13) and the widely used isothermal titration calorimetry (ITC) (14, 15). As with NMR, ITC has many advantages, but exhibits modest sensitivity and often requires large sample quantities. Another increasingly popular free-solution approach is microscale thermophoresis (...

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Section: Introductionmentioning

confidence: 75%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Origin and prediction of free-solution interaction studies performed label-free

Bornhop

Kammer

Kussrow

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…Sota et.al. (28) used molar ellipticity of the sample molecule in combination with surface plasmon resonance (SPR), also an RI detector, to show a relation between surface and bulk signals. The underlying implication of their observations may be that an RI detector could be used to measure binding events in free solution.…”

mentioning

confidence: 99%

Free-Solution, Label-Free Molecular Interactions Studied by Back-Scattering Interferometry

Bornhop

Latham

Kussrow

et al. 2007

Science

191

216

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Free-solution, label-free molecular interactions were investigated with back-scattering interferometry in a simple optical train composed of a helium-neon laser, a microfluidic channel, and a position sensor. Molecular binding interactions between proteins, ions and protein, and small molecules and protein, were determined with high dynamic range dissociation constants (K d spanning six decades) and unmatched sensitivity (picomolar K d 's and detection limits of 10,000s of molecules). With this technique, equilibrium dissociation constants were quantified for protein A and immunoglobulin G, interleukin-2 with its monoclonal antibody, and calmodulin with calcium ion Ca 2+ , a small molecule inhibitor, the protein calcineurin, and the M13 peptide. The high sensitivity of back-scattering interferometry and small volumes of microfluidics allowed the entire calmodulin assay to be performed with 200 picomoles of solute.

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“…Some methods have been used to improve the sensitivity of SPR sensor for detecting small molecules. For instance, employed polymers with high molecular weight as a transduction layer to increase the changes in the refractive index at the sensing interface triggered by small molecules [91], utilized the conformational change of macromolecules or proteins switched by small molecules to amplify the change of SPR signal [92,93], used of a high molecular weight ligand that competed with the small molecular weight analyte for receptor binding. Liu [94] detailed the incorporation of a water-soluble deep cavitand into a membrane bilayer assembled onto a nanoglassified surface for study of molecular recognition in a membrane-mimicking setting.…”

Section: The Development Of Spr Sensor and Spr Commercial Instrumentsmentioning

confidence: 99%

Development and Application of Time-Resolved Surface Plasmon Resonance Spectrometer

Mao¹,

Bao²,

Wang³

et al. 2011

AJAC

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Surface plasmon resonance (SPR) sensor, an optical sensor exploiting special electromagnetic waves-surface plasmon polaritons to probe interactions between an analyte in solution and a molecular recognition element immobilized on the SPR sensor surface, has been widely used in various realms, such as investigating biomolecular interactions and binding properties, detection of biological and chemical analytes, environmental monitoring, food safety and medical diagnostics. This paper reviews the development of SPR sensors and SPR commercial instruments, and emphatically introduces the time-resolved surface plasmon resonance (TR-SPR) techniques. The excellent performances of high sensitivity and rapid detection are easily achieved with TR-SPR spectrometer, whereas the traditional SPR spectrometer cannot be accomplished. Therefore, TR-SPR spectrometer is appropriate for real time analysis of bio-recognition events and small molecular dynamics. However, only two commercial TR-SPR instruments have been exhibited at present, Thermo FT-SPR 100 and Autolab ESPRIT (SPRINGLE). Unfortunately, the high-priced instruments are not suitable for extensive applications in laboratories at present. Herein, a novel commercial TR-SPR spectrometer has been introduced in this review.

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Detection of Conformational Changes in an Immobilized Protein Using Surface Plasmon Resonance

Cited by 170 publications

References 29 publications

Origin and prediction of free-solution interaction studies performed label-free

Origin and prediction of free-solution interaction studies performed label-free

Free-Solution, Label-Free Molecular Interactions Studied by Back-Scattering Interferometry

Development and Application of Time-Resolved Surface Plasmon Resonance Spectrometer

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