New tool for spreading proteins to the environment: Cry1Ab toxin immobilized to bioplastics

Moldes, Cristina; Farinós, Gema P.; Eugenio, Laura I. de; Garcı́a, Pedro; Garcı́a, José Luis; Ortego, Félix; Hernández‐Crespo, Pedro; Castañera, Pedro; Prieto, María Auxiliadora

doi:10.1007/s00253-005-0257-6

Cited by 18 publications

(26 citation statements)

References 27 publications

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“…This disorder-order transition is typical of most of the intrinsic disordered proteins described so far [62]. Our results may help to understand the function of these important proteins involved in the metabolism of the PHA bioplastics, and could also be of utility to obtain improved variants of the BioF affinity tag [27], [28]. Moreover, some structural features of PhaF are shared among a high number of other phasins, even from unrelated organisms, namely the conservation of coiled-coil oligomerization sequences and an appreciable level of intrinsic disorder, evidences that deserve further investigations.…”

Section: Discussionsupporting

confidence: 52%

“…However, the neat separation into domains that can be discerned from the inspection of its sequence [17] and experimental evidence [18], [27], [28] (see Introduction) prompted us to model such domains separately, and subsequently link the resulting structures. Then, the model was subjected to experimental validation using hydrodynamic and spectroscopical techniques.…”

Section: Discussionmentioning

confidence: 99%

“…Also, the N-terminal domain is conserved in other PHA-binding phasins with a different C-terminal part [17], [18]. Moreover, very diverse fusion proteins containing the N-terminal moiety of PhaF (the BioF affinity tag) can also be adsorbed on polyhydroxyalkanoate granules without compromising their function [27], [28]. Finally, the isolated C-terminal moiety is able to bind to DNA [18].…”

Section: Introductionmentioning

confidence: 99%

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A New Family of Intrinsically Disordered Proteins: Structural Characterization of the Major Phasin PhaF from Pseudomonas putida KT2440

et al. 2013

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Phasins are intracellular polyhydroxyalkanoat4e (PHA)-associated proteins involved in the stabilization of these bacterial carbon storage granules. Despite its importance in PHA metabolism and regulation, only few reports have focused so far on the structure of these proteins. In this work we have investigated the structure and stability of the PhaF phasin from Pseudomonas putida KT2440, a protein that is involved in PHA granule stabilization and distribution to daughter cells upon cell division. A structural, three-dimensional model of the protein was built from homology modeling procedures and consensus secondary structure predictions. The model predicts that PhaF is an elongated protein, with a long, amphipathic N-terminal helix with PHA binding capacity, followed by a short leucine zipper involved in protein oligomerization and a superhelical C-terminal domain wrapped around the chromosomal DNA. Hydrodynamic, spectroscopical and thermodynamic experiments validated the model and confirmed both that free PhaF is a tetramer in solution and that most part of the protein is intrinsically disordered in the absence of its ligands. The results lay a molecular basis for the explanation of the biological role of PhaF and, along with an exhaustive analysis of phasin sequence databases, suggest that intrinsic disorder and oligomerization through coiled-coils may be a widespread mechanism among these proteins.

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Section: Discussionsupporting

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A New Family of Intrinsically Disordered Proteins: Structural Characterization of the Major Phasin PhaF from Pseudomonas putida KT2440

et al. 2013

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“…These granules could be easily purified by a simple centrifugation step and proteins subsequently released by treatment with mild detergents, maintaining their enzymatic activity (52). This method was proposed for the release of proteins to the environment, in particular for the liberation of the Cry1Ab toxin as an insecticide treatment (53). misfolded proteins appear in the cell as a result of different kinds of stress, such as heat shock, production of PHB, or overproduction of heterologous proteins, chaperones bind to them, and RpoH freed from chaperones activates the heat shock response.…”

Section: Biotechnological Applications Of Phasinsmentioning

confidence: 99%

Phasins, Multifaceted Polyhydroxyalkanoate Granule-Associated Proteins

Mezzina

Pettinari

2016

Appl Environ Microbiol

107

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Phasins are the major polyhydroxyalkanoate (PHA) granule-associated proteins. They promote bacterial growth and PHA synthesis and affect the number, size, and distribution of the granules. These proteins can be classified in 4 families with distinctive characteristics. Low-resolution structural studies and in silico predictions were performed in order to elucidate the structure of different phasins. Most of these proteins share some common structural features, such as a preponderant ␣-helix composition, the presence of disordered regions that provide flexibility to the protein, and coiled-coil interacting regions that form oligomerization domains. Due to their amphiphilic nature, these proteins play an important structural function, forming an interphase between the hydrophobic content of PHA granules and the hydrophilic cytoplasm content. Phasins have been observed to affect both PHA accumulation and utilization. Apart from their role as granule structural proteins, phasins have a remarkable variety of additional functions. Different phasins have been determined to (i) activate PHA depolymerization, (ii) increase the expression and activity of PHA synthases, (iii) participate in PHA granule segregation, and (iv) have both in vivo and in vitro chaperone activities. These properties suggest that phasins might play an active role in PHA-related stress protection and fitness enhancement. Due to their granule binding capacity and structural flexibility, several biotechnological applications have been developed using different phasins, increasing the interest in the study of these remarkable proteins.

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“…A simple expression of a fusion tag comprising the Cry toxin protein fused to the N-terminal of BioF P.pu allowed for the fabrication of granules[49]. A few years later, researchers coined a the desired recombinant proteins are immobilized/displayed in a single phase of bacterial cells, i.e.…”

mentioning

confidence: 99%

Recombinant surface engineering to enhance and expand the potential of biologically produced nanoparticles: A review

Kutralam-Muniasamy

Pérez-Guevara

2017

Process Biochemistry

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New tool for spreading proteins to the environment: Cry1Ab toxin immobilized to bioplastics

Cited by 18 publications

References 27 publications

A New Family of Intrinsically Disordered Proteins: Structural Characterization of the Major Phasin PhaF from Pseudomonas putida KT2440

A New Family of Intrinsically Disordered Proteins: Structural Characterization of the Major Phasin PhaF from Pseudomonas putida KT2440

Phasins, Multifaceted Polyhydroxyalkanoate Granule-Associated Proteins

Recombinant surface engineering to enhance and expand the potential of biologically produced nanoparticles: A review

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