scholarly journals Combinatorial design of chemical-dependent protein switches for controlling intracellular electron transfer

2019 ◽  
Author(s):  
Bingyan Wu ◽  
Joshua T. Atkinson ◽  
Dimithree Kahanda ◽  
George. N. Bennett ◽  
Jonathan J. Silberg
Keyword(s):  
Electron Transfer ◽  
Ligand Binding ◽  
Fusion Proteins ◽  
Electron Flow ◽  
Combinatorial Design ◽  
Human Estrogen Receptor ◽  
Dependent Protein ◽  
Insertion Sites ◽  
Domain Insertion ◽  
Bacterial Selection

ABSTRACTOne challenge with controlling electron flow in cells is the lack of biomolecules that directly couple the sensing of environmental conditions to electron transfer efficiency. To overcome this protein component limitation, we randomly inserted the ligand binding domain (LBD) from the human estrogen receptor (ER) into a thermostable 2Fe-2S ferredoxin (Fd) from Mastigocladus laminosus and used a bacterial selection to identify Fd-LBD fusion proteins that support electron transfer from a Fd-NADP reductase (FNR) to a Fd-dependent sulfite reductase (SIR). Mapping LBD insertion sites onto structure revealed that Fd tolerates domain insertion adjacent to or within the tetracysteine motif that coordinates the 2Fe-2S metallocluster. With both classes of the fusion proteins, cellular ET was enhanced by the ER antagonist 4-hydroxytamoxifen. In addition, one of Fds arising from ER-LBD insertion within the tetracysteine motif acquires an oxygen-tolerant 2Fe-2S cluster, suggesting that ET is regulated through post-translational ligand binding.

scholarly journals Combinatorial design of chemical‐dependent protein switches for controlling intracellular electron transfer

AIChE Journal ◽  
2019 ◽  
Vol 66 (3) ◽  
Author(s):  
Bingyan Wu ◽  
Joshua T. Atkinson ◽  
Dimithree Kahanda ◽  
George N. Bennett ◽  
Jonathan J. Silberg
Keyword(s):  
Electron Transfer ◽  
Combinatorial Design ◽  
Dependent Protein

Hypothesis: versatile function of ferredoxin-NADP+ reductase in cyanobacteria provides regulation for transient photosystem I-driven cyclic electron flow

2002 ◽  
Vol 29 (3) ◽  
pp. 201 ◽  
Author(s):  
Hans C. P. Matthijs ◽  
Robert Jeanjean ◽  
Nataliya Yeremenko ◽  
Jef Huisman ◽  
Francoise Joset ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Salt Stress ◽  
De Novo ◽  
Electron Flow ◽  
High Co2 ◽  
Partial Restoration ◽  
Synechocystis Pcc6803 ◽  
Low Co2 ◽  
Cyclic Electron Transfer

Pseudo-reversion of the high-CO2 requiring phenotype of the NADH dehydrogenase type 1-impaired mutant of Synechocystis PCC6803, strain M55, by salt stress coincides with partial restoration of PSI-driven cyclic electron transfer. In M55, the complete family of D proteins (D1–D6) that are needed for electron transfer through the complex is lacking. Adaptation to salt stress requires de novo synthesis of full-length 47-kDa ferredoxin-NADP+ reductase (FNR). A mutant created in the M55 background, which only expresses truncated chloroplast 37-kDa FNR cannot adapt to salt stress and refrains from growth in low CO2. A special feature of FNR in cyanobacteria is the relatively high molecular mass of 44–48 kDa. A positively charged extended N-terminal domain of the cyanobacterial enzyme defines the extra mass. The extension likely plays a key role in the salt-stress inducible enhancement of PSI-driven cyclic electron transfer, and in the pseudo-reversion of the high-CO2 requiring phenotype of M55. Data acquired with several other cyanobacteria and the oxychlorobacterium Prochlorothrix hollandica contributed to the present hypothesis. It proposes that FNR is involved in regulation of inducible and transient PSI cyclic electron transfer in cyanobacteria via a thylakoid surface charge and conditional-proteolysis steered mechanism.

scholarly journals Converting a Periplasmic Binding Protein into a Synthetic Biosensing Switch through Domain Insertion

2019 ◽  
Vol 2019 ◽  
pp. 1-15 ◽  
Author(s):  
Lucas F. Ribeiro ◽  
Vanesa Amarelle ◽  
Liliane F. C. Ribeiro ◽  
María-Eugenia Guazzaroni
Keyword(s):  
Protein Engineering ◽  
Ligand Binding ◽  
Conformational Changes ◽  
Protein Function ◽  
Cellular Response ◽  
Protein Domain ◽  
Signal Molecule ◽  
Synthetic Protein ◽  
Periplasmic Binding Proteins ◽  
Domain Insertion

All biosensing platforms rest on two pillars: specific biochemical recognition of a particular analyte and transduction of that recognition into a readily detectable signal. Most existing biosensing technologies utilize proteins that passively bind to their analytes and therefore require wasteful washing steps, specialized reagents, and expensive instruments for detection. To overcome these limitations, protein engineering strategies have been applied to develop new classes of protein-based sensor/actuators, known as protein switches, responding to small molecules. Protein switches change their active state (output) in response to a binding event or physical signal (input) and therefore show a tremendous potential to work as a biosensor. Synthetic protein switches can be created by the fusion between two genes, one coding for a sensor protein (input domain) and the other coding for an actuator protein (output domain) by domain insertion. The binding of a signal molecule to the engineered protein will switch the protein function from an “off” to an “on” state (or vice versa) as desired. The molecular switch could, for example, sense the presence of a metabolite, pollutant, or a biomarker and trigger a cellular response. The potential sensing and response capabilities are enormous; however, the recognition repertoire of natural switches is limited. Thereby, bioengineers have been struggling to expand the toolkit of molecular switches recognition repertoire utilizing periplasmic binding proteins (PBPs) as protein-sensing components. PBPs are a superfamily of bacterial proteins that provide interesting features to engineer biosensors, for instance, immense ligand-binding diversity and high affinity, and undergo large conformational changes in response to ligand binding. The development of these protein switches has yielded insights into the design of protein-based biosensors, particularly in the area of allosteric domain fusions. Here, recent protein engineering approaches for expanding the versatility of protein switches are reviewed, with an emphasis on studies that used PBPs to generate novel switches through protein domain insertion.

Characterization of ligand binding and electron-transfer properties of Lucina pectinata hemoglobin I

1995 ◽  
Vol 59 (2-3) ◽  
pp. 447
Author(s):  
Manuel Maldonado ◽  
Angela M. Navarro ◽  
Juan González-Lagoa ◽  
Juan López-Garriga ◽  
Jorge L. Colón
Keyword(s):  
Electron Transfer ◽  
Ligand Binding ◽  
Lucina Pectinata ◽  
Electron Transfer Properties ◽  
Transfer Properties

scholarly journals Electric Field Induced Biomimetic Transmembrane Electron Transport using Carbon Nanotube Porins as Bipolar Electrodes

2021 ◽  
Author(s):  
Jacqueline M. Hicks ◽  
Yun-Chiao Yao ◽  
Sydney Barber ◽  
Aleksandr Noy ◽  
Nigel Neate ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Carbon Nanotube ◽  
Electron Transport ◽  
Electric Fields ◽  
Electron Flow ◽  
Transport Systems ◽  
Redox Systems ◽  
Bipolar Electrodes ◽  
Electrochemical Approach ◽  
Transmembrane Electron Transport

<p>Cells modulate their homeostasis through the control of redox reactions via transmembrane electron transport systems. These are largely mediated via oxidoreductase enzymes. Their use in biology has been linked to a host of systems including reprogramming for energy requirements in cancer. Consequently, our ability to modulate membrane redox systems may give rise to opportunities to modulate underlying biology. The current work aimed to develop a wireless bipolar electrochemical approach to form on-demand electron transfer across biological membranes. To achieve this goal, we show that using membrane inserted carbon nanotube porins that can act as bipolar nanoelectrodes, we could control electron flow with externally applied electric fields across membranes. Before this work, bipolar electrochemistry has been thought to require high applied voltages not compatible with biological systems. We show that bipolar electrochemical reaction via gold reduction at the nanotubes could be modulated at low cell-friendly voltages, providing an opportunity to use bipolar electrodes to control electron flux across membranes. Our observations present a new opportunity to use bipolar electrodes to alter cell behavior via wireless control of membrane electron transfer.</p>

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