Biomod/2013/NanoUANL/Background

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<strong><span style="font-size:24px;"><span style="font-family: tahoma,geneva,sans-serif;">BACKGROUND</span></span></strong></p>
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<strong><span style="font-size:24px;"><span style="font-family: tahoma,geneva,sans-serif;">&nbsp;&nbsp; BACKGROUND</span></span></strong></p>
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<span style="font-size:14px;"><span style="font-family: lucida sans unicode,lucida grande,sans-serif;"><strong><img alt="" src="http://openwetware.org/images/9/97/UANLBackground1.jpg" style="width: 635px; height: 401px; float: center;" /></strong></span></span></p>
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<span style="font-size:14px;"><span style="font-family: lucida sans unicode,lucida grande,sans-serif;"><strong><img alt="" src="http://openwetware.org/images/9/97/UANLBackground1.jpg" style="width: 635px; height: 375px;" /></strong></span></span></p>
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&nbsp;</p>
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<span style="font-size:14px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;"><strong>Reactions in Containers</strong></span></span></p>
<span style="font-size:14px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;"><strong>Reactions in Containers</strong></span></span></p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;">The most widely studied containers are liposomes: self-assembling structures formed by a lipid bilayer, that have been used to encapsulate enzymes for a variety of applications. &nbsp;However, other biological systems have also been subject of study, such as block copolymer vesicles, proteins like ferritin, and viral capsids.</span></span></p>
<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;">The most widely studied containers are liposomes: self-assembling structures formed by a lipid bilayer, that have been used to encapsulate enzymes for a variety of applications. &nbsp;However, other biological systems have also been subject of study, such as block copolymer vesicles, proteins like ferritin, and viral capsids.</span></span></p>
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&nbsp;</p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;"><img alt="" src="http://openwetware.org/images/6/65/UANLBackground2.jpg" style="width: 463px; height: 383px;" /></span></span></p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;"><span style="font-size:11px;">Fig 1. Different viral structures, both with and without a viral envelope</span></span></span></p>
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&nbsp;</p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;">Viral capsids have gained the interest of researchers for some of their properties. The mechanical stability that this structure presents is similar to that of other structures, which protects the container&rsquo;s cargo. Many viral containers have been used as templates for nanoparticle synthesis.</span></span></p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;">The CCMV virus has been the subject of study for nanotechnology for its unique properties, even among viruses. The container can be disassembled and reassembled at will, depending on the environmental conditions and pores allow the entrance or exit of small molecules.</span><span style="font-size:12px;"> To explore more properties, the capsid has been modified in a variety of ways, allowing us to change the N-terminal amino region.</span></span></p>
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<span style="font-size:14px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;"><strong>Enzyme (HRP)</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;">Horseradish peroxidase (HRP) is a ~44 kDa glycoprotein from the peroxidase family, with a known three-dimensional structure. Peroxidases typically catalyze a reaction in which a wide variety of both organic and inorganic compounds are oxidized.</span></span></p>
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<span style="font-size:12px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;">There has been a great deal of scientific interest in HRP because of its commercial uses, primarily as a component of clinical diagnostic kits and for immunostaining. The enzyme is usually conjugated to specific antibodies or streptavidin, which binds to the compound of interest and activity, is detected with substrates like TMB or ABTS, which are then analyzed by a colorimetric assay. Enzymatic reactions inside CCMV have been studied as single-molecule system, using HRP as the catalyst. The researchers showed that the interior of the capsid presented a suitable environment for enzymatic activity. The reaction was examined by fluorescence microscopy using a fluorogenic substrate.</span></span></p>
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&nbsp;</p>
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<span style="font-size:14px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;"><strong>Capsid-assisted Synthesis</strong></span></span></p>
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<span style="font-size:12px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;">The biomimetic approach to material synthesis consists in the combination of well-defined structures, macromolecular templates and molecular interactions. Soluble precursors can be transformed into nanoparticles by a variety of mechanisms inside a viral capsid, such as pH changes. Results have suggested as hypothesis that the condensation of certain materials inside containers is electrostatically driven, and the selection of the metal precursor influences this process.</span></span><br />
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<span style="font-size:12px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;">An advantage to these processes is the minimization of hazardous substances used and generated by these reactions. Additionally, these substances are often expensive and need special containment, which is problematic. The reduction of metal ions by combinations of biomolecules found in the extracts of certain organisms (e.g., enzymes/proteins, amino acids, polysaccharides, and vitamins) is environmentally friendly, yet chemically complex, making this system fascinating to study. Silver nanoparticle synthesis has been reported in bacteria, fungi and plants.</span></span></p>
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<span _fck_bookmark="1" style="display: none;">&nbsp;</span><img alt="" src="http://openwetware.org/images/7/74/UANLBackground3.jpg" style="width: 350px; height: 349px;" /><span _fck_bookmark="1" style="display: none;">&nbsp;</span></p>
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<span style="font-size:11px;"><span style="font-family: trebuchet ms,helvetica,sans-serif;">Fig. 2 - 3D structure of the complete CCMV capsid</span></span></p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;">As a nanoreactor, the CCMV capsid is more versatile than other viruses, since its reversible pH-dependent disassembly/assembly behavior permits the encapsulation of enzymes and metal nanoparticles. However, it has fewer examples of functionalization because of the lower thermodynamic stability of the structure, compared to other viruses.</span></span></p>
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<span style="font-family:trebuchet ms,helvetica,sans-serif;"><span style="font-size: 12px;">CCMV has been previously used to form insoluble TiO2 nanoparticles from soluble Ti(IV) salts inside the capsid. A modified CCMV structure with a negative charge has also been used to selectively bind Fe(II) and Fe(III) to the internal surface. Selective mineralization with the TMV capsid has resulted in Au(0) mineralization on the viral surface, and Ag(0) mineralization within its interior. CoPt and FePt nanowires have been fabricated by incorporating specific nucleating peptides on the surface of the M13 virus coat structure.</span></span></p>
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Revision as of 13:50, 26 October 2013

UANL_Banner2.png

   BACKGROUND

 

 

Reactions in Containers

Nanocontainer is a term used for structures with a size within the nanometer range (1-100 nm). The interest in their use stems from their empty inner cavities that can be used for a variety of applications. Among these, one of peculiar interest is the encapsulation of molecules in order to turn the structure into a reaction vessel.

Biological systems have always been an inspiration because of their complexity and diversity.  Cell processes take place within constrained spaces and small volumes, and many are not yet fully understood. Advances in nanoscale fabrication have allowed us to mimic some of these spaces and features with other structures. The volumes that are present at this level (atto and zeptoliter) allow molecules to collide more often, as opposed to an “open space”; simulations based on Brownian diffusion have shown that collision frequency between molecules strongly depend on vesicle size.

The most widely studied containers are liposomes: self-assembling structures formed by a lipid bilayer, that have been used to encapsulate enzymes for a variety of applications.  However, other biological systems have also been subject of study, such as block copolymer vesicles, proteins like ferritin, and viral capsids.

 

Fig 1. Different viral structures, both with and without a viral envelope

 

Viral capsids have gained the interest of researchers for some of their properties. The mechanical stability that this structure presents is similar to that of other structures, which protects the container’s cargo. Many viral containers have been used as templates for nanoparticle synthesis.

The CCMV virus has been the subject of study for nanotechnology for its unique properties, even among viruses. The container can be disassembled and reassembled at will, depending on the environmental conditions and pores allow the entrance or exit of small molecules. To explore more properties, the capsid has been modified in a variety of ways, allowing us to change the N-terminal amino region.

 

Enzyme (HRP)

Horseradish peroxidase (HRP) is a ~44 kDa glycoprotein from the peroxidase family, with a known three-dimensional structure. Peroxidases typically catalyze a reaction in which a wide variety of both organic and inorganic compounds are oxidized.

There has been a great deal of scientific interest in HRP because of its commercial uses, primarily as a component of clinical diagnostic kits and for immunostaining. The enzyme is usually conjugated to specific antibodies or streptavidin, which binds to the compound of interest and activity, is detected with substrates like TMB or ABTS, which are then analyzed by a colorimetric assay. Enzymatic reactions inside CCMV have been studied as single-molecule system, using HRP as the catalyst. The researchers showed that the interior of the capsid presented a suitable environment for enzymatic activity. The reaction was examined by fluorescence microscopy using a fluorogenic substrate.

 

Capsid-assisted Synthesis

The biomimetic approach to material synthesis consists in the combination of well-defined structures, macromolecular templates and molecular interactions. Soluble precursors can be transformed into nanoparticles by a variety of mechanisms inside a viral capsid, such as pH changes. Results have suggested as hypothesis that the condensation of certain materials inside containers is electrostatically driven, and the selection of the metal precursor influences this process.
An advantage to these processes is the minimization of hazardous substances used and generated by these reactions. Additionally, these substances are often expensive and need special containment, which is problematic. The reduction of metal ions by combinations of biomolecules found in the extracts of certain organisms (e.g., enzymes/proteins, amino acids, polysaccharides, and vitamins) is environmentally friendly, yet chemically complex, making this system fascinating to study. Silver nanoparticle synthesis has been reported in bacteria, fungi and plants.


  

Fig. 2 - 3D structure of the complete CCMV capsid

As a nanoreactor, the CCMV capsid is more versatile than other viruses, since its reversible pH-dependent disassembly/assembly behavior permits the encapsulation of enzymes and metal nanoparticles. However, it has fewer examples of functionalization because of the lower thermodynamic stability of the structure, compared to other viruses.


CCMV has been previously used to form insoluble TiO2 nanoparticles from soluble Ti(IV) salts inside the capsid. A modified CCMV structure with a negative charge has also been used to selectively bind Fe(II) and Fe(III) to the internal surface. Selective mineralization with the TMV capsid has resulted in Au(0) mineralization on the viral surface, and Ag(0) mineralization within its interior. CoPt and FePt nanowires have been fabricated by incorporating specific nucleating peptides on the surface of the M13 virus coat structure.

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