A comparative study of recombinant and native frutalin binding to human prostate tissues

Oliveira, Carla Cristina Marques de; Teixeira, J. A.; Schmitt, Fernando; Domingues, Lucı́lia

doi:10.1186/1472-6750-9-78

Cited by 19 publications

(15 citation statements)

References 30 publications

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“…Figure 2 illustrates the schematic pathway from protein production to purification with the studied solubility tags (His 6 tag, GST, MBP, NusA, Trx, SUMO, H, and Fh8). Here, the selected target proteins included the 12-kDa surface protein of C. parvum (CP12), the lectin frutalin from the Artocarpus incisa plant (FTL; Oliveira et al, 2008 , 2009a , b , 2011 ), and four proteins from the yeast S. cerevisiae : reduced viability upon starvation protein 167 (RVS167), phospholipase D1 (SPO14), and serine/threonine-protein kinases 1 and 2 (YPK1 and YPK2). These target proteins were all known as difficult-to-express in E. coli , and presented different molecular weights, locations, and functions.…”

Section: The Novel Fh8 Fusion System (Hitag®)mentioning

confidence: 99%

Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

et al. 2014

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Proteins are now widely produced in diverse microbial cell factories. The Escherichia coli is still the dominant host for recombinant protein production but, as a bacterial cell, it also has its issues: the aggregation of foreign proteins into insoluble inclusion bodies is perhaps the main limiting factor of the E. coli expression system. Conversely, E. coli benefits of cost, ease of use and scale make it essential to design new approaches directed for improved recombinant protein production in this host cell. With the aid of genetic and protein engineering novel tailored-made strategies can be designed to suit user or process requirements. Gene fusion technology has been widely used for the improvement of soluble protein production and/or purification in E. coli, and for increasing peptide’s immunogenicity as well. New fusion partners are constantly emerging and complementing the traditional solutions, as for instance, the Fh8 fusion tag that has been recently studied and ranked among the best solubility enhancer partners. In this review, we provide an overview of current strategies to improve recombinant protein production in E. coli, including the key factors for successful protein production, highlighting soluble protein production, and a comprehensive summary of the latest available and traditionally used gene fusion technologies. A special emphasis is given to the recently discovered Fh8 fusion system that can be used for soluble protein production, purification, and immunogenicity in E. coli. The number of existing fusion tags will probably increase in the next few years, and efforts should be taken to better understand how fusion tags act in E. coli. This knowledge will undoubtedly drive the development of new tailored-made tools for protein production in this bacterial system.

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Section: The Novel Fh8 Fusion System (Hitag®)mentioning

confidence: 99%

Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

et al. 2014

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“…To overcome these limitations, frutalin with a defined amino acid sequence was expressed and produced in the Pichia pastoris expression system [ 15 ]. The ability of this recombinant frutalin and native frutalin to bind human tumour cells has recently been demonstrated [ 16 , 17 ]. Moreover, recombinant frutalin showed higher capacity than native frutalin to differentiate malignant from nonmalignant cells in immunohistochemical studies conducted with human prostate tissues [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

Cytotoxic Effects of Native and Recombinant Frutalin, a Plant Galactose‐Binding Lectin, on HeLa Cervical Cancer Cells

Oliveira

Nicolau

Teixeira

et al. 2011

BioMed Research International

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Frutalin is the α-D-galactose-binding lectin isolated from breadfruit seeds. Frutalin was obtained from two different sources: native frutalin was purified from its natural origin, and recombinant frutalin was produced and purified from Pichia pastoris. This work aimed to study and compare the effect of native and recombinant frutalin on HeLa cervical cancer cells proliferation and apoptosis. Furthermore, the interaction between frutalin and the HeLa cells was investigated by confocal microscopy. Despite having different carbohydrate-binding affinities, native and recombinant frutalin showed an identical magnitude of cytotoxicity on HeLa cells growth (IC50~100 μg/mL) and equally induced cell apoptosis. The interaction studies showed that both lectins were rapidly internalised and targeted to HeLa cell's nucleus. Altogether, these results indicate that frutalin action is not dependent on its sugar-binding properties. This study provides important information about the bioactivity of frutalin and contributes to the understanding of the plant lectins cytotoxic activity.

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“…The potential of recombinant and native frutalin, α -D-galactose-binding plant lectin from Artocarpus incisa seeds to marker human prostate tumor, was evaluated [ 158 ]. Prostate carcinoma and benign prostate hyperplasia tissues were analyzed using the lectins conjugated with anti-frutalin polyclonal antibody to react with a complex biotinylated anti-rabbit IgG and streptavidin-conjugated peroxidase, followed by the addition of DAB for visualization of lectin staining.…”

Section: Lectins As Histochemical Markersmentioning

confidence: 99%

Lectins, Interconnecting Proteins with Biotechnological/Pharmacological and Therapeutic Applications

Coelho

Silva

Lima

et al. 2017

Evidence-Based Complementary and Alternative Medicine

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Lectins are proteins extensively used in biomedical applications with property to recognize carbohydrates through carbohydrate-binding sites, which identify glycans attached to cell surfaces, glycoconjugates, or free sugars, detecting abnormal cells and biomarkers related to diseases. These lectin abilities promoted interesting results in experimental treatments of immunological diseases, wounds, and cancer. Lectins obtained from virus, microorganisms, algae, animals, and plants were reported as modulators and tool markers in vivo and in vitro; these molecules also play a role in the induction of mitosis and immune responses, contributing for resolution of infections and inflammations. Lectins revealed healing effect through induction of reepithelialization and cicatrization of wounds. Some lectins have been efficient agents against virus, fungi, bacteria, and helminths at low concentrations. Lectin-mediated bioadhesion has been an interesting characteristic for development of drug delivery systems. Lectin histochemistry and lectin-based biosensors are useful to detect transformed tissues and biomarkers related to disease occurrence; antitumor lectins reported are promising for cancer therapy. Here, we address lectins from distinct sources with some biological effect and biotechnological potential in the diagnosis and therapeutic of diseases, highlighting many advances in this growing field.

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A comparative study of recombinant and native frutalin binding to human prostate tissues

Cited by 19 publications

References 30 publications

Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

Cytotoxic Effects of Native and Recombinant Frutalin, a Plant Galactose‐Binding Lectin, on HeLa Cervical Cancer Cells

Lectins, Interconnecting Proteins with Biotechnological/Pharmacological and Therapeutic Applications

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