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Selection of peptides binding to metallic borides by screening M13 phage display libraries.

Ploss M, Facey SJ, Bruhn C, Zemel L, Hofmann K, Stark RW, Albert B, Hauer B - BMC Biotechnol. (2014)

Bottom Line: The 7-mer peptide sequence LGFREKE, isolated on amorphous Ni3B emerged as the best binder for both substrates.Fluorescence microscopy and atomic force microscopy confirmed the specific binding affinity of LGFREKE expressing phage to amorphous and crystalline Ni3B nanoparticles.We think that the identified strong binding sequences described here could potentially serve for the utilisation of M13 phage as a viable alternative to other methods to create tailor-made boride composite materials or new catalytic surfaces by a biologically driven nano-assembly synthesis and structuring.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. bernhard.hauer@itb.uni-stuttgart.de.

ABSTRACT

Background: Metal borides are a class of inorganic solids that is much less known and investigated than for example metal oxides or intermetallics. At the same time it is a highly versatile and interesting class of compounds in terms of physical and chemical properties, like semiconductivity, ferromagnetism, or catalytic activity. This makes these substances attractive for the generation of new materials. Very little is known about the interaction between organic materials and borides. To generate nanostructured and composite materials which consist of metal borides and organic modifiers it is necessary to develop new synthetic strategies. Phage peptide display libraries are commonly used to select peptides that bind specifically to metals, metal oxides, and semiconductors. Further, these binding peptides can serve as templates to control the nucleation and growth of inorganic nanoparticles. Additionally, the combination of two different binding motifs into a single bifunctional phage could be useful for the generation of new composite materials.

Results: In this study, we have identified a unique set of sequences that bind to amorphous and crystalline nickel boride (Ni3B) nanoparticles, from a random peptide library using the phage display technique. Using this technique, strong binders were identified that are selective for nickel boride. Sequence analysis of the peptides revealed that the sequences exhibit similar, yet subtle different patterns of amino acid usage. Although a predominant binding motif was not observed, certain charged amino acids emerged as essential in specific binding to both substrates. The 7-mer peptide sequence LGFREKE, isolated on amorphous Ni3B emerged as the best binder for both substrates. Fluorescence microscopy and atomic force microscopy confirmed the specific binding affinity of LGFREKE expressing phage to amorphous and crystalline Ni3B nanoparticles.

Conclusions: This study is, to our knowledge, the first to identify peptides that bind specifically to amorphous and to crystalline Ni3B nanoparticles. We think that the identified strong binding sequences described here could potentially serve for the utilisation of M13 phage as a viable alternative to other methods to create tailor-made boride composite materials or new catalytic surfaces by a biologically driven nano-assembly synthesis and structuring.

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Analysis of the phage binding by fluorescence microscopy. Confocal fluorescence (A, C) and transmission optical microscopy (B, D) of LGFREKE phage bound to amorphous (A, B) and crystalline (C, D) Ni3B nanoparticles. The samples were dyed using a fluorescently tagged anti-M13 monoclonal antibody. Confocal fluorescence (E, G) and transmission optical microscopy (F, H) of the control experiments of amorphous and crystalline Ni3B nanoparticles, respectively, which were preincubated with M13KE wild-type (M13wt) phage before incubation with the fluorescently tagged antibody.
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Figure 3: Analysis of the phage binding by fluorescence microscopy. Confocal fluorescence (A, C) and transmission optical microscopy (B, D) of LGFREKE phage bound to amorphous (A, B) and crystalline (C, D) Ni3B nanoparticles. The samples were dyed using a fluorescently tagged anti-M13 monoclonal antibody. Confocal fluorescence (E, G) and transmission optical microscopy (F, H) of the control experiments of amorphous and crystalline Ni3B nanoparticles, respectively, which were preincubated with M13KE wild-type (M13wt) phage before incubation with the fluorescently tagged antibody.

Mentions: The binding of phage displaying the A7 peptide LGFREKE to amorphous and crystalline Ni3B nanoparticles was confirmed by fluorescence microscopy. Amplified phage displaying the LGFREKE peptide were incubated either with amorphous or crystalline Ni3B nanoparticles and then washed to remove the nonspecifically bound phage particles. The samples were then dyed using a fluorescently tagged anti-M13 monoclonal antibody and washed again to eliminate nonspecifically bound anti-M13 antibodies. The fluorescence confocal images in Figure 3A and C show the LGFREKE displaying phage binding to the surface of amorphous and crystalline Ni3B nanoparticles, respectively. Considerable fluorescence emits from the surfaces of the LGFREKE phage-Ni3B nanoparticles. Although the relative binding affinity experiments showed an unspecific binding of the wild-type phage to the Ni3B substrates, the control experiments (Figure 3E and G) showed no fluorescence. This could be due to the fact that the concentration of bound phage is below the detection limit.


Selection of peptides binding to metallic borides by screening M13 phage display libraries.

Ploss M, Facey SJ, Bruhn C, Zemel L, Hofmann K, Stark RW, Albert B, Hauer B - BMC Biotechnol. (2014)

Analysis of the phage binding by fluorescence microscopy. Confocal fluorescence (A, C) and transmission optical microscopy (B, D) of LGFREKE phage bound to amorphous (A, B) and crystalline (C, D) Ni3B nanoparticles. The samples were dyed using a fluorescently tagged anti-M13 monoclonal antibody. Confocal fluorescence (E, G) and transmission optical microscopy (F, H) of the control experiments of amorphous and crystalline Ni3B nanoparticles, respectively, which were preincubated with M13KE wild-type (M13wt) phage before incubation with the fluorescently tagged antibody.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3924706&req=5

Figure 3: Analysis of the phage binding by fluorescence microscopy. Confocal fluorescence (A, C) and transmission optical microscopy (B, D) of LGFREKE phage bound to amorphous (A, B) and crystalline (C, D) Ni3B nanoparticles. The samples were dyed using a fluorescently tagged anti-M13 monoclonal antibody. Confocal fluorescence (E, G) and transmission optical microscopy (F, H) of the control experiments of amorphous and crystalline Ni3B nanoparticles, respectively, which were preincubated with M13KE wild-type (M13wt) phage before incubation with the fluorescently tagged antibody.
Mentions: The binding of phage displaying the A7 peptide LGFREKE to amorphous and crystalline Ni3B nanoparticles was confirmed by fluorescence microscopy. Amplified phage displaying the LGFREKE peptide were incubated either with amorphous or crystalline Ni3B nanoparticles and then washed to remove the nonspecifically bound phage particles. The samples were then dyed using a fluorescently tagged anti-M13 monoclonal antibody and washed again to eliminate nonspecifically bound anti-M13 antibodies. The fluorescence confocal images in Figure 3A and C show the LGFREKE displaying phage binding to the surface of amorphous and crystalline Ni3B nanoparticles, respectively. Considerable fluorescence emits from the surfaces of the LGFREKE phage-Ni3B nanoparticles. Although the relative binding affinity experiments showed an unspecific binding of the wild-type phage to the Ni3B substrates, the control experiments (Figure 3E and G) showed no fluorescence. This could be due to the fact that the concentration of bound phage is below the detection limit.

Bottom Line: The 7-mer peptide sequence LGFREKE, isolated on amorphous Ni3B emerged as the best binder for both substrates.Fluorescence microscopy and atomic force microscopy confirmed the specific binding affinity of LGFREKE expressing phage to amorphous and crystalline Ni3B nanoparticles.We think that the identified strong binding sequences described here could potentially serve for the utilisation of M13 phage as a viable alternative to other methods to create tailor-made boride composite materials or new catalytic surfaces by a biologically driven nano-assembly synthesis and structuring.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. bernhard.hauer@itb.uni-stuttgart.de.

ABSTRACT

Background: Metal borides are a class of inorganic solids that is much less known and investigated than for example metal oxides or intermetallics. At the same time it is a highly versatile and interesting class of compounds in terms of physical and chemical properties, like semiconductivity, ferromagnetism, or catalytic activity. This makes these substances attractive for the generation of new materials. Very little is known about the interaction between organic materials and borides. To generate nanostructured and composite materials which consist of metal borides and organic modifiers it is necessary to develop new synthetic strategies. Phage peptide display libraries are commonly used to select peptides that bind specifically to metals, metal oxides, and semiconductors. Further, these binding peptides can serve as templates to control the nucleation and growth of inorganic nanoparticles. Additionally, the combination of two different binding motifs into a single bifunctional phage could be useful for the generation of new composite materials.

Results: In this study, we have identified a unique set of sequences that bind to amorphous and crystalline nickel boride (Ni3B) nanoparticles, from a random peptide library using the phage display technique. Using this technique, strong binders were identified that are selective for nickel boride. Sequence analysis of the peptides revealed that the sequences exhibit similar, yet subtle different patterns of amino acid usage. Although a predominant binding motif was not observed, certain charged amino acids emerged as essential in specific binding to both substrates. The 7-mer peptide sequence LGFREKE, isolated on amorphous Ni3B emerged as the best binder for both substrates. Fluorescence microscopy and atomic force microscopy confirmed the specific binding affinity of LGFREKE expressing phage to amorphous and crystalline Ni3B nanoparticles.

Conclusions: This study is, to our knowledge, the first to identify peptides that bind specifically to amorphous and to crystalline Ni3B nanoparticles. We think that the identified strong binding sequences described here could potentially serve for the utilisation of M13 phage as a viable alternative to other methods to create tailor-made boride composite materials or new catalytic surfaces by a biologically driven nano-assembly synthesis and structuring.

Show MeSH
Related in: MedlinePlus