Dimerization of a flocculent protein from<i>Moringa oleifera</i>: experimental evidence and<i>in silico</i>interpretation

Pavankumar, Asalapuram R.; Kayathri, Rajarathinam; Murugan, N.; Zhang, Qiong; Srivastava, Vaibhav; Okoli, Chuka; Bulone, Vincent; Rajarao, Gunaratna Kuttuva; Ågren, Hans

doi:10.1080/07391102.2013.770374

Cited by 13 publications

(12 citation statements)

References 28 publications

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“…The nonpolar contribution to the desolvation free energy is computed using the solvent accessible surface area. The polar contribution is computed using either generalised Born approach (in GB approach) or Poisson Boltzmann approach (as in PB approach)434445. Overall, these two approaches differ with respect to the polar term in the desolvation free energy.…”

Section: Resultsmentioning

confidence: 99%

On/off-switchable LSPR nano-immunoassay for troponin-T

Ashaduzzaman

Deshpande

Murugan

et al. 2017

Sci Rep

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Regeneration of immunosensors is a longstanding challenge. We have developed a re-usable troponin-T (TnT) immunoassay based on localised surface plasmon resonance (LSPR) at gold nanorods (GNR). Thermosensitive poly(N-isopropylacrylamide) (PNIPAAM) was functionalised with anti-TnT to control the affinity interaction with TnT. The LSPR was extremely sensitive to the dielectric constant of the surrounding medium as modulated by antigen binding after 20 min incubation at 37 °C. Computational modelling incorporating molecular docking, molecular dynamics and free energy calculations was used to elucidate the interactions between the various subsystems namely, IgG-antibody (c.f., anti-TnT), PNIPAAM and/or TnT. This study demonstrates a remarkable temperature dependent immuno-interaction due to changes in the PNIPAAM secondary structures, i.e., globular and coil, at above or below the lower critical solution temperature (LCST). A series of concentrations of TnT were measured by correlating the λLSPR shift with relative changes in extinction intensity at the distinct plasmonic maximum (i.e., 832 nm). The magnitude of the red shift in λLSPR was nearly linear with increasing concentration of TnT, over the range 7.6 × 10−15 to 9.1 × 10−4 g/mL. The LSPR based nano-immunoassay could be simply regenerated by switching the polymer conformation and creating a gradient of microenvironments between the two states with a modest change in temperature.

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Section: Resultsmentioning

confidence: 99%

On/off-switchable LSPR nano-immunoassay for troponin-T

Ashaduzzaman

Deshpande

Murugan

et al. 2017

Sci Rep

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“…Indeed, both the hemagglutinin MoL and the flocculating lectin cMoL presented molecular profiles similar to Mo -CBP 2 , appearing in dimer or trimer conformations (Katre et al, 2008; Luz et al, 2013). The in silico analysis of the recombinant protein MO 2.1 showed that the dimer form was the most stable structural arrangement (Pavankumar et al, 2014). Thus, it is plausible to suggest that the M. oleifera proteins aggregate into oligomers to attain a stable and lower energy structural state.…”

Section: Discussionmentioning

confidence: 99%

A Chitin-binding Protein Purified from Moringa oleifera Seeds Presents Anticandidal Activity by Increasing Cell Membrane Permeability and Reactive Oxygen Species Production

et al. 2017

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Candida species are opportunistic pathogens that infect immunocompromised and/or immunosuppressed patients, particularly in hospital facilities, that besides representing a significant threat to health increase the risk of mortality. Apart from echinocandins and triazoles, which are well tolerated, most of the antifungal drugs used for candidiasis treatment can cause side effects and lead to the development of resistant strains. A promising alternative to the conventional treatments is the use of plant proteins. M. oleifera Lam. is a plant with valuable medicinal properties, including antimicrobial activity. This work aimed to purify a chitin-binding protein from M. oleifera seeds and to evaluate its antifungal properties against Candida species. The purified protein, named Mo-CBP2, represented about 0.2% of the total seed protein and appeared as a single band on native PAGE. By mass spectrometry, Mo-CBP2 presented 13,309 Da. However, by SDS-PAGE, Mo-CBP2 migrated as a single band with an apparent molecular mass of 23,400 Da. Tricine-SDS-PAGE of Mo-CBP2 under reduced conditions revealed two protein bands with apparent molecular masses of 7,900 and 4,600 Da. Altogether, these results suggest that Mo-CBP2 exists in different oligomeric forms. Moreover, Mo-CBP2 is a basic glycoprotein (pI 10.9) with 4.1% (m/m) sugar and it did not display hemagglutinating and hemolytic activities upon rabbit and human erythrocytes. A comparative analysis of the sequence of triptic peptides from Mo-CBP2 in solution, after LC-ESI-MS/MS, revealed similarity with other M. oleifera proteins, as the 2S albumin Mo-CBP3 and flocculating proteins, and 2S albumins from different species. Mo-CBP2 possesses in vitro antifungal activity against Candida albicans, C. parapsilosis, C. krusei, and C. tropicalis, with MIC50 and MIC90 values ranging between 9.45–37.90 and 155.84–260.29 μM, respectively. In addition, Mo-CBP2 (18.90 μM) increased the cell membrane permeabilization and reactive oxygen species production in C. albicans and promoted degradation of circular plasmid DNA (pUC18) from Escherichia coli. The data presented in this study highlight the potential use of Mo-CBP2 as an anticandidal agent, based on its ability to inhibit Candida spp. growth with apparently low toxicity on mammalian cells.

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“…Crude seed extract (MOCE) was prepared by grinding the seeds followed by solvent treatments as detailed in Ghebremicahel et al The purified fractions (1 µg µL −1 ) of MOCP and MOCRP were obtained from our earlier study . All three protein fractions (MOCE, MOCP and MOCRP) were resolved on 10% SDS‐PAGE stained with Coomassie Brilliant Blue‐R250.…”

Section: Methodsmentioning

confidence: 99%

“…The molecular docking studies were performed using the homology‐modeled structure of MOCP, recently reported by us, which was docked to establish a stable complex with congo red, tartrazine and DDT to analyze their affinity and molecular interactions. The standardized parameters of the molecular docking tool, AutoDock 4.2 was used to compute the molecular interactions between the protein and ligands (pollutants).…”

Section: Methodsmentioning

confidence: 99%

“…Among the crude seed proteins, a 6.5 kDa water‐soluble protein is widely reported for its applications in water treatment. We have recently reported that this 6.5 kDa oligomeric protein (MO coagulant protein; MOCP) exhibits very good antimicrobial and coagulant activities, either in its natural (obtained from seed) or recombinant form . Considering the importance of MOCP, we have produced it recombinantly and scaled‐up its production using bioreactors for large‐scale water treatment applications and also demonstrated its coagulation activity with 0.01 mg L −1 and antimicrobial response using as low as 10 mg L −1 .…”

Section: Introductionmentioning

confidence: 99%

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Identification ofMoringa oleiferaprotein responsible for the decolorization and pesticide removal from drinking water and industrial effluent - anin silicoandin situevaluation

Pavankumar

Singh

2014

J. Chem. Technol. Biotechnol.

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Natural products are always in demand, especially in the food and water treatment industry, to reduce health hazards caused by the prolonged use of chemicals. Though crude seed extract of Moringa oleifera (MOCE) is used for decolouration, the protein responsible for such activity is not fully known. In this study, in silico analysis of Moringa oleifera coagulant protein (MOCP; a predominant oligomeric protein in MOCE) was undertaken to check its molecular interactions with water and soil pollutants, in order to identify the protein accountable for such activities. The molecular docking studies of MOCP with azo dyes like congo red, tartrazine) and a pesticide (dichlorodiphenyltrichloroethane) revealed a strong binding affinity (−5.66, −5.33 and −5.04, respectively, kJ mol−1) between the protein and the pollutants through hydrogen bonding and electrostatic interactions. Further, these results were verified in situ with MOCP, a recombinant form of MOCP (MOCRP) and MOCE against congo red (100 mg L−1) and revealed the dye removal efficiency of 63.8%, 65.7%, and 72.3%, respectively. While the jar test results of synthetic coloured water and industrial textile effluent containing congo red showed 51.6% and 58.3%. Hence, we believe that the MOCP is responsible for multiple activities of MOCE and suggest its prospective use for large‐scale treatment of drinking water and industrial effluents. © 2014 Society of Chemical Industry

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Dimerization of a flocculent protein fromMoringa oleifera: experimental evidence andin silicointerpretation

Cited by 13 publications

References 28 publications

On/off-switchable LSPR nano-immunoassay for troponin-T

On/off-switchable LSPR nano-immunoassay for troponin-T

A Chitin-binding Protein Purified from Moringa oleifera Seeds Presents Anticandidal Activity by Increasing Cell Membrane Permeability and Reactive Oxygen Species Production

Identification ofMoringa oleiferaprotein responsible for the decolorization and pesticide removal from drinking water and industrial effluent - anin silicoandin situevaluation

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