Enzyme adsorption-induced activity changes: a quantitative study on TiO2 model agglomerates

Márquez, Andrés; Kocsis, Krisztina; Zickler, Gregor A.; Bourret, Gilles R.; Feinle, Andrea; Hüsing, Nicola; Himly, Martin; Duschl, Albert; Berger, Thomas; Diwald, Oliver

doi:10.1186/s12951-017-0283-4

Cited by 16 publications

(10 citation statements)

References 49 publications

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“…Data analysis was performed with the program Origin (OriginLab Corporation, Northampton, MA, USA). To estimate the content of secondary structure, the amide I band in the spectra of the protein in D 2 O was deconvoluted following a procedure described previously [ 62 ]. Briefly, the second derivative of the experimental spectra (upon subtraction of the straight region over the range 1800–2000 cm −1 ) was used to identify the number and position of the overlapping components bands, whose single contribution to the experimental curves was obtained by the iterative Chi square minimization of the combination of Gaussian-type peaks using the Levenberg-Marquardt algorithm.…”

Section: Methodsmentioning

confidence: 99%

The Recombinant Inhibitor of DNA Binding Id2 Forms Multimeric Structures via the Helix-Loop-Helix Domain and the Nuclear Export Signal

Roschger

Schubert

Regl

et al. 2018

IJMS

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The inhibitor of DNA binding and cell differentiation 2 (Id2) is a helix-loop-helix (HLH) protein that acts as negative dominant regulator of basic-HLH transcription factors during development and in cancer. The structural properties of Id2 have been investigated so far by using synthetic or recombinant fragments reproducing single domains (N-terminus, HLH, C-terminus): the HLH domain tends to dimerize into a four-helix bundle, whereas the flanking regions are flexible. In this work, the intact protein was expressed in E. coli, solubilized from inclusion bodies with urea, purified and dissolved in water at pH~4. Under these conditions, Id2 was obtained with both cysteine residues disulfide-bonded to β-mercaptoethanol that was present during the solubilization process. Moreover, it existed in a self-assembled state, in which the N-terminus remained highly flexible, while the HLH domain and, surprisingly, part of the C-terminus, which corresponds to the nuclear export signal (NES), both were involved in slowly tumbling, rigid structures. The protein oligomers also formed twisted fibrils that were several micrometers long and up to 80 nm thick. These results show that self-assembly decreases the backbone flexibility of those two protein regions (HLH and NES) that are important for interaction with basic-HLH transcription factors or for nucleocytoplasmic shuttling.

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Section: Methodsmentioning

confidence: 99%

The Recombinant Inhibitor of DNA Binding Id2 Forms Multimeric Structures via the Helix-Loop-Helix Domain and the Nuclear Export Signal

Roschger

Schubert

Regl

et al. 2018

IJMS

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“…Such conformational changes have been induced mainly by adsorptive interactions 33,34 . The main target of both approaches has been, as expected, the active site 32,34,35 , but far less attention has been devoted to affecting the channel leading to the active site. The rationale in approaching the (dynamic) channel is that it acts like a highly stereoselective separation column, allowing entrance to the active site only for a substrate that ts the stereoselective screening.…”

Section: Discussionmentioning

confidence: 92%

Glucose oxidase converted into a general sugar-oxidase

Baruch-Shpigler

Avnir

2022

Preprint

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Entrapment of glucose oxidase (GOx) within metallic gold converts this widely used enzyme into a general saccharide oxidase. The following sugar molecules were oxidized by the entrapped enzyme (in addition to D-glucose): L-glucose, fructose, xylose,, glucose-6-phosphate, sucrose, lactose, methylglucoside, and the tri-saccharide raffinose. None of these sugars except raffinose has a natural specific oxidase. The origin of this generalization of activity is attributed to the strong protein-gold 3D interactions and to the strong interactions of co-entrapped cetyltrimethylammonium bromide (CTAB) with gold on one hand, and with the protein on the other hand. It is proposed that these interactions induce conformational changes in the channel leading to the active site, which is located at the interface between the two units of the dimeric GOx protein. The observations are compatible with affecting the specific conformation change of pulling apart and opening this gate-keeper, rendering the active site accessible to a variety of substrates. In addition to converting a common enzyme to an oxidase for common sugars which lack it, the entrapment methodology was found to be beneficial in increasing the thermal stability of GOx up to 100 oC and in allowing its convenient recyclability, two features of practical importance.

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“…This secondary structure is difficult to analyse due to spectral overlapping and band's intensity. Hence, complementary techniques used to extract “hidden” information inside a specific vibration band, such as second‐derivative and/or deconvolution, are applied during the processing of the vibrational spectra …”

Section: Introductionmentioning

confidence: 88%

A comprehensive study of protein–mesoporous–macroporous silica interactions by extended canonical variate analysis of Raman spectra

Videira‐Quintela

Prazeres

Ortega-Ojeda

et al. 2019

J Raman Spectroscopy

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Understanding the protein–support interactions is of major importance when manufacturing bionanomaterials to a certain application. These interactions can be the cause for enhanced properties or denaturation phenomena in the target protein. Raman spectroscopy was applied to a bionanomaterial comprehending the protein β‐galactosidase immobilized by physical adsorption into a mesoporous–macroporous silica material, with a nanoporous network consisting of 9‐nm mesopores and 200‐nm macropores. Raman spectra of the bionanomaterial evidenced a complex amount of differences related to the Raman shifts, intensities, band enlargement, appearance of new bands, and overlapping, in comparison with the silica support and the protein spectra. To help in the analysis of the Raman spectra and in the inspection of possible protein–support interactions, ECVA (extended canonical variate analysis) was used as a chemometric complementary tool, dividing the spectra into four segments: 1 (3,100 to 2,800 cm−1), 2 (1,800 to 1,500 cm−1), 3 (1,500 to 1,200 cm−1), and 4 (1,200 to 900 cm−1). Major alterations in the Amide I band (1,800 to 1,500 cm−1) and the amino acid band regions demonstrated possible structure alterations to a non‐native form of the protein β‐galactosidase. Also, other minor alterations were observed in other spectral regions (3,100 to 2,800 cm−1,1,500 to 1,200 cm−1, and 1,200 to 900 cm−1) also representative of protein structure alteration due to protein–support interactions.

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Enzyme adsorption-induced activity changes: a quantitative study on TiO2 model agglomerates

Cited by 16 publications

References 49 publications

The Recombinant Inhibitor of DNA Binding Id2 Forms Multimeric Structures via the Helix-Loop-Helix Domain and the Nuclear Export Signal

The Recombinant Inhibitor of DNA Binding Id2 Forms Multimeric Structures via the Helix-Loop-Helix Domain and the Nuclear Export Signal

Glucose oxidase converted into a general sugar-oxidase

A comprehensive study of protein–mesoporous–macroporous silica interactions by extended canonical variate analysis of Raman spectra

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