Role of histidine 148 in stability and dynamics of a highly fluorescent GFP variant

Campanini, Barbara; Pioselli, Barbara; Raboni, Samanta; Felici, Paolo; Giordano, I.; D’Alfonso, Laura; Collini, Maddalena; Chirico, Giuseppe; Bettati, Stefano

doi:10.1016/j.bbapap.2013.01.014

Cited by 10 publications

(9 citation statements)

References 53 publications

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“…39,40 Thus, it represents an excellent target to assess how ncAA can be used to influence critical interactions that define function. Replacement of H148 with azF results in a ∼90 nm blue shift in the major absorbance peak to the less favourable 400 nm (Fig.…”

Section: Resultsmentioning

confidence: 99%

Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle

et al. 2016

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Section: Resultsmentioning

confidence: 99%

Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle

et al. 2016

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“…In fact, as shown in Figure 7, after four cycles of reuse the percentage of activity is almost the same either if the puuE-doped silica gel is rinsed with a potassium phosphate buffer containing 150 mM NaCl or if the salt is absent. As previously observed for other encapsulated proteins [51][52][53][54], the role of counter-ions in shielding the excess negative charges exposed on the surface of gel pores at near physiological pH is important to avoid partitioning of polar solutes and aspecific protein-pore interactions, a possible source of functional heterogeneity for entrapped molecules. In this case, electrostatic interactions between the pores and charged groups on protein surface seem not to limit functionally relevant dynamics.…”

Section: Reusability Of the Biosensor And Stability Over Timementioning

confidence: 72%

Immobilization of Allantoinase for the Development of an Optical Biosensor of Oxidative Stress States

Marchetti

Ronda

Percudani

et al. 2019

Sensors

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Allantoin, the natural end product of purine catabolism in mammals, is non-enzymatically produced from the scavenging of reactive oxygen species through the degradation of uric acid. Levels of allantoin in biological fluids are sensitively influenced by the presence of free radicals, making this molecule a candidate marker of acute oxidative stress in clinical analyses. With this aim, we exploited allantoinase-the enzyme responsible for allantoin hydrolization in plants and lower organisms-for the development of a biosensor exploiting a fast enzymatic-chemical assay for allantoin quantification. Recombinant allantoinase was entrapped in a wet nanoporous silica gel matrix and its structural properties, function, and stability were characterized through fluorescence spectroscopy and circular dichroism measurements, and compared to the soluble enzyme. Physical immobilization in silica gel minimally influences the structure and the catalytic efficiency of entrapped allantoinase, which can be reused several times and stored for several months with good activity retention. These results, together with the relative ease of the sol-gel preparation and handling, make the encapsulated allantoinase a good candidate for the development of an allantoin biosensor.

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“…Overall, data indicate that the encapsulated β-galactosidase is less stable than the enzyme in solution, likely due to alteration of the conformational distribution. A decreased stability for an encapsulated enzyme with respect to the soluble form is unusual as the gel matrix tends to prevent unfolding, which is accompanied by volume expansion [63,64].…”

Section: Enzyme Encapsulation In Silica Gels For Protein Delivery Bimentioning

confidence: 99%

More than a Confinement: “Soft” and “Hard” Enzyme Entrapment Modulates Biological Catalyst Function

et al. 2019

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Catalysis makes chemical and biochemical reactions kinetically accessible. From a technological point of view, organic, inorganic, and biochemical catalysis is relevant for several applications, from industrial synthesis to biomedical, material, and food sciences. A heterogeneous catalyst, i.e., a catalyst confined in a different phase with respect to the reagents’ phase, requires either its physical confinement in an immobilization matrix or its physical adsorption on a surface. In this review, we will focus on the immobilization of biological catalysts, i.e., enzymes, by comparing hard and soft immobilization matrices and their effect on the modulation of the catalysts’ function. Indeed, unlike smaller molecules, the catalytic activity of protein catalysts depends on their structure, conformation, local environment, and dynamics, properties that can be strongly affected by the immobilization matrices, which, therefore, not only provide physical confinement, but also modulate catalysis.

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Role of histidine 148 in stability and dynamics of a highly fluorescent GFP variant

Cited by 10 publications

References 53 publications

Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle

Molecular basis for functional switching of GFP by two disparate non-native post-translational modifications of a phenyl azide reaction handle

Immobilization of Allantoinase for the Development of an Optical Biosensor of Oxidative Stress States

More than a Confinement: “Soft” and “Hard” Enzyme Entrapment Modulates Biological Catalyst Function

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