SDS-PAGE and IR spectroscopy to evaluate modifications in the viral protein profile induced by a cationic porphyrinic photosensitizer

Costa, Liliana; Esteves, Ana Cristina; Correia, António; Moreirinha, Catarina; Delgadillo, Ivonne; Neves, Maria G. P. M. S.; Faustino, Maria A. F.; Almeida, Adelaide

doi:10.1016/j.jviromet.2014.09.013

Cited by 17 publications

(17 citation statements)

References 51 publications

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“…Photodynamic inactivation (PDI) of viruses has been shown to be an efficient alternative to antiviral agents in the control of resistant and emerging viruses [1][2][3][4][5][6][7][8][9][10]. When irradiated with visible light and in the presence of molecular oxygen in aqueous solution, photosensitisers such as 5,10,15,20-tetrakis (1-methyl-4-pyridinio) porphyrin tetra p-toluenesulfonate (TMPyP) can generate singlet oxygen by a type 2 reaction (and other reactive oxygen species (ROS) by type 1 reaction) [11][12][13]. Singlet oxygen and other ROS can react rapidly and cause irreversible damage to biomolecules thereby leading to the inactivation of viruses and other microorganisms [11,[14][15][16][17][18][19].…”

Section: Introductionmentioning

confidence: 99%

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Photodynamic inactivation of bacteriophage MS2: The A-protein is the target of virus inactivation

Majiya

Adeyemi

Stonehouse

et al. 2018

Journal of Photochemistry and Photobiology B: Biology

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Singlet oxygen mediated oxidation has been shown to be responsible for photodynamic inactivation (PDI) of viruses in solution with photosensitisers such as 5, 10, 15, 20-tetrakis (1-methyl-4-pyridinio) porphyrin tetra p-toluenesulfonate (TMPyP). The capsids of non-enveloped viruses, such as bacteriophage MS2, are possible targets for viral inactivation by singlet oxygen oxidation. Within the capsid (predominantly composed of coat protein), the A-protein acts as the host recognition and attachment protein. The A-protein has two domains; an α-helix domain and a β-sheet domain. The α-helix domain is attached to the viral RNA genome inside the capsid while the β-sheet domain, which is on the surface of the capsid, is believed to be the site for attachment to the host bacteria pilus during infection. In this study, 4 sequence-specific antibodies were raised against 4 sites on the A-protein. Changes induced by the oxidation of singlet oxygen were compared to the rate of PDI of the virus. Using these antibodies, our results suggest that the rate of PDI is relative to loss of antigenicity of two sites on the A-protein. Our data further showed that PDI caused aggregation of MS2 particles and crosslinking of MS2 coat protein. However, these inter- and intra-capsid changes did not correlate to the rate of PDI we observed in MS2. Possible modes of action are discussed as a means to gaining insight to the targets and mechanisms of PDI of viruses.

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Section: Introductionmentioning

confidence: 99%

“…Changes to viral proteins as result of PDI have been measured by SDS-PAGE and infrared (IR) spectroscopy [12]. However, these methods are not specific to PDI-mediated effects.…”

Section: Introductionmentioning

confidence: 99%

Photodynamic inactivation of bacteriophage MS2: The A-protein is the target of virus inactivation

Majiya

Adeyemi

Stonehouse

et al. 2018

Journal of Photochemistry and Photobiology B: Biology

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“…Recent work on the PDI of viruses with porphyrins has focused on cationic porphyrin derivatives, specifically N-methylpyridyl-substituted compounds. 75,89,98,200,201,202,203,204,205,206,207,208 Cationic porphyrins are well known to be effective in the PDI of bacteria as well as viruses. 5,15,209,210,211,212 Costa et al investigated the structure-activity relationship in a series of cationic porphyrins carrying two to four charges in the PDI of the T4-like sewage phage, a non-enveloped double stranded DNA virus.…”

Section: Porphyrinsmentioning

confidence: 99%

“…Selection of structures used in the QSAR study on the antiviral activity of Nmethylpyridyl-substituted porphyrins by Costa et al 202a Costa et al have subsequently used compounds 20 and 21 in more detail in several related publications, 75,89,203,204,205 using the T4like sewage phage as a model, as well as other phages. PDI of phage T4 with these porphyrins was shown to be dependent on the light source, fluence rate and total light dose with phage activation being more effective at lower fluence rates.…”

Section: Figure 10mentioning

confidence: 99%

Trends and targets in antiviral phototherapy

Wiehe

O’Brien

Senge

2019

Photochem Photobiol Sci

219

168

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“…46,47 The separation was performed in the Mini-PROTEAN 3 (Bio-Rad) with lab casted SDS polyacrylamide gels (15%). Gels ran for 2 h, at 120 V and at 4°C.…”

Section: Protein Electrophoresis (Sds-page)mentioning

confidence: 99%

Protein profiles of Escherichia coli and Staphylococcus warneri are altered by photosensitization with cationic porphyrins

Alves

Esteves

Correia

et al. 2015

Photochem Photobiol Sci

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Oxidative stress induced by photodynamic treatment of microbial cells causes irreversible damages to vital cellular components such as proteins. Photodynamic inactivation (PDI) of bacteria, a promising therapeutic approach for the treatment of superficial and localized skin and oral infections, can be achieved by exciting a photosensitizing agent with visible light in an oxygenated environment. Although some studies have addressed the oxidative alterations of PDI in bacterial proteins, the present study is the first to compare the electrophoretic profiles of proteins of Gram-positive and Gram-negative bacteria, having two structurally different porphyrins, with different kinetics of photoinactivation. The cationic porphyrins 5,10,15-tris(1-methylpyridinium-4-yl)-20-(pentafluorophenyl)porphyrin tri-iodide (Tri-Py(+)-Me-PF) and 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)porphyrin tetra-iodide (Tetra-Py(+)-Me) were used to photosensitize Escherichia coli and Staphylococcus warneri upon white light irradiation at an irradiance of 4.0 mW cm(-2). After different photosensitization periods, proteins were extracted from bacteria and analyzed using one-dimensional SDS-PAGE. Apparent molecular weights and band intensities were determined after an irradiation period corresponding to a reduction of 4 log10 in cell viability. After photodynamic treatment, there was a general loss of bacterial proteins, assigned to large-scale protein degradation. Protein loss was more pronounced after PDI with Tri-Py(+)-Me-PF in both bacteria. There was also an increase in the concentration of some proteins as well as an increase in the molecular weight of other proteins. We show that proteins of E. coli and S. warneri are important targets of PDI. Although there is an attempt of cellular response to the PDI-induced damage by overexpression of a limited number of proteins, the damage is lethal. Our results show that changes occurring in the protein pattern during photodynamic treatment are different with the two photosensitizers, which helps to explain the different inactivation kinetics of the two bacteria. SDS-PAGE is a rational approach to assign the type of cellular response to stress that is being induced in the cells.

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SDS-PAGE and IR spectroscopy to evaluate modifications in the viral protein profile induced by a cationic porphyrinic photosensitizer

Cited by 17 publications

References 51 publications

Photodynamic inactivation of bacteriophage MS2: The A-protein is the target of virus inactivation

Photodynamic inactivation of bacteriophage MS2: The A-protein is the target of virus inactivation

Trends and targets in antiviral phototherapy

Protein profiles of Escherichia coli and Staphylococcus warneri are altered by photosensitization with cationic porphyrins

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