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'''Bifunctional Proteinaceous Hydrogels for Bioelectrocatalysis'''  
'''Engineering of Conformational Changes in Peptides'''  


Self-assembly is an essential process for all forms of life. For example, proteins spontaneously fold into well-defined 3-dimensional structures, and cellular organelles form that spatially segregate diverse cellular processes.  As engineers aim to create  new devices and systems at ever decreasing size scales, self-assembly processes become increaseingly attactive techniques.
Proteins are involved in almost every aspect of cellular function, and therefore they have evolved with a myriad of properties and capabilities. The catalytic nature of proteins is widely appreciated, and the field of Protein Engineering has emerged over the past 25 years to both study and improve the biochemical properties of proteins for a wide variety of important applications.


We are collaborating with Plamen Atanassov at University of New Mexico, Scott Calabrese-Barton at Michigan State University and Shelley Minteer at Saint Louis University to make improved electrodes for biofuel cells.  In a biofuel cell, redox enzymes are immobilized on electrode surfaces.  Enzymes located at the anode are able to oxidize substrates and the electrons move through an external circuit to create power.  On the cathode, other redox enzymes are able to use the electrons to reduce oxygen to water.  The net result of this process is the generation of electricity from a variety of readily available biofuel sources, with oxygen as the terminal electron acceptor.
More recently, the physical properties and abilities of proteins and peptides have been gaining attention. For example, the hemagglutinin protein from the influenza virus undergoes a dramatic pH-triggered conformational change to promote membrane fusion during infection. 
 
As our understanding of the natural conformational plasticity of proteins and peptides expands, these properties are increasingly being exploited for the design and engineering of new protein and peptide-based systems.  For example, the well-known elastin-like peptides are known to undergo a dramatic reversible conformational change, from a disordered peptide to a collapsed b-spiral with reduced solubility, in response to changes in temperature, pH, or ionic strength.  This transition has been termed the inverse temperature transition, due to the fact that the peptides become more ordered (not less) with increased temperature, and this effect has been extensively characterized.  The elastin-like peptide conformational change has been used as a building-block for a variety of rationally engineered systems, including advanced bioseparations, environmental remediation, drug delivery, molecular switches, tissue engineering, and bionanotechnology.
 
We are developing a new methods for the Directed Evolution of peptides that exhibit novel conformational behavior in response to environmental stimuli.  We are developing new sensing systems that can be used to report the conformational behavior of an attached peptide.  One approach is to use a single chain antibody fragment (scFv) that has a variable binding affinity depending on the conformational state of its linker peptide.  The new peptides that can be discovered using these creative approaches will be extensively characterized, which will to help to further elucidate the molecular underpinnings of the conformational behavior of peptides.  And, the new peptides will be immediately valuable for use in many avenues of research including biotechnology, synthetic biology, nanotechnology, biosensing and drug discovery.


The architecture of the electrodes is crucial for biofuel cell performance.  The enzymes on the electrodes must be positioned so that electrons can easily move between the enzymatic active site and the electorde surface (Direct Electron Transfer (DET)).  Alternatively, the enzymes can be immoblilized with redox mediators, such as osmium, that facilitate the transport of electrons from the electrodes to the enzymes (Fig. 1).  In this Mediated Electron Transport (MET) configuration, the enzyme and mediators are immobilized in a polymer matrix on the electrode surface.  While this system has been used to demonstrate impressive biofuel cell performances, it is potentially hampered by poor dispersion of the enzyme and mediator within the polymer matrix, and complex manufacturing requirments.


We are using biological self-assembly to improve the biofuel cell electrode construction and performance (Fig. 2).  Instead of combing enzymes and mediators in a polymer matrix, we are creating self-assembling protein-based hydrogels that intrinsically include the redox enzymes and the mediators.  In this configuration the loading of the enzyme and the mediators into the hydrogel can be finely controlled, and the hydrogel assembly process will be well-defined and repeatable.  These new bioelectrocatalytic hydrogels will have the potential to significantly improve biofuel cell performance.




'''Related Publications'''
'''Related Publications'''


<biblio>
<biblio>
 
#Paper6 pmid=20420519
#Paper5 pmid=19454225
#Paper4 pmid=18411225
#Paper3 pmid=18218715
#Paper2 pmid=17376876
#Paper1 pmid=17450770
</biblio>
</biblio>

Latest revision as of 14:02, 9 December 2011

Banta Lab

Protein and Metabolic Engineering

Home Lab Members Publications Research Interests Courses Pictures Positions Available


Engineering of Conformational Changes in Peptides

Proteins are involved in almost every aspect of cellular function, and therefore they have evolved with a myriad of properties and capabilities. The catalytic nature of proteins is widely appreciated, and the field of Protein Engineering has emerged over the past 25 years to both study and improve the biochemical properties of proteins for a wide variety of important applications.

More recently, the physical properties and abilities of proteins and peptides have been gaining attention. For example, the hemagglutinin protein from the influenza virus undergoes a dramatic pH-triggered conformational change to promote membrane fusion during infection.

As our understanding of the natural conformational plasticity of proteins and peptides expands, these properties are increasingly being exploited for the design and engineering of new protein and peptide-based systems. For example, the well-known elastin-like peptides are known to undergo a dramatic reversible conformational change, from a disordered peptide to a collapsed b-spiral with reduced solubility, in response to changes in temperature, pH, or ionic strength. This transition has been termed the inverse temperature transition, due to the fact that the peptides become more ordered (not less) with increased temperature, and this effect has been extensively characterized. The elastin-like peptide conformational change has been used as a building-block for a variety of rationally engineered systems, including advanced bioseparations, environmental remediation, drug delivery, molecular switches, tissue engineering, and bionanotechnology.

We are developing a new methods for the Directed Evolution of peptides that exhibit novel conformational behavior in response to environmental stimuli. We are developing new sensing systems that can be used to report the conformational behavior of an attached peptide. One approach is to use a single chain antibody fragment (scFv) that has a variable binding affinity depending on the conformational state of its linker peptide. The new peptides that can be discovered using these creative approaches will be extensively characterized, which will to help to further elucidate the molecular underpinnings of the conformational behavior of peptides. And, the new peptides will be immediately valuable for use in many avenues of research including biotechnology, synthetic biology, nanotechnology, biosensing and drug discovery.



Related Publications

  1. Banta S, Wheeldon IR, and Blenner M. Protein engineering in the development of functional hydrogels. Annu Rev Biomed Eng. 2010 Aug 15;12:167-86. DOI:10.1146/annurev-bioeng-070909-105334 | PubMed ID:20420519 | HubMed [Paper6]
  2. Chockalingam K, Lu HD, and Banta S. Development of a bacteriophage-based system for the selection of structured peptides. Anal Biochem. 2009 May 1;388(1):122-7. DOI:10.1016/j.ab.2009.01.042 | PubMed ID:19454225 | HubMed [Paper5]
  3. Casali M, Banta S, Zambonelli C, Megeed Z, and Yarmush ML. Site-directed mutagenesis of the hinge peptide from the hemagglutinin protein: enhancement of the pH-responsive conformational change. Protein Eng Des Sel. 2008 Jun;21(6):395-404. DOI:10.1093/protein/gzn018 | PubMed ID:18411225 | HubMed [Paper4]
  4. Blenner MA and Banta S. Characterization of the 4D5Flu single-chain antibody with a stimulus-responsive elastin-like peptide linker: a potential reporter of peptide linker conformation. Protein Sci. 2008 Mar;17(3):527-36. DOI:10.1110/ps.073257308 | PubMed ID:18218715 | HubMed [Paper3]
  5. Chockalingam K, Blenner M, and Banta S. Design and application of stimulus-responsive peptide systems. Protein Eng Des Sel. 2007 Apr;20(4):155-61. DOI:10.1093/protein/gzm008 | PubMed ID:17376876 | HubMed [Paper2]
  6. Banta S, Megeed Z, Casali M, Rege K, and Yarmush ML. Engineering protein and peptide building blocks for nanotechnology. J Nanosci Nanotechnol. 2007 Feb;7(2):387-401. DOI:10.1166/jnn.2007.153 | PubMed ID:17450770 | HubMed [Paper1]

All Medline abstracts: PubMed | HubMed

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