scholarly journals Chromatophores efficiently promote light-driven ATP synthesis and DNA transcription inside hybrid multicompartment artificial cells

2021 ◽  
Vol 118 (7) ◽  
pp. e2012170118
Author(s):  
Emiliano Altamura ◽  
Paola Albanese ◽  
Roberto Marotta ◽  
Francesco Milano ◽  
Michele Fiore ◽  
...  

The construction of energetically autonomous artificial protocells is one of the most ambitious goals in bottom-up synthetic biology. Here, we show an efficient manner to build adenosine 5′-triphosphate (ATP) synthesizing hybrid multicompartment protocells. Bacterial chromatophores from Rhodobacter sphaeroides accomplish the photophosphorylation of adenosine 5′-diphosphate (ADP) to ATP, functioning as nanosized photosynthetic organellae when encapsulated inside artificial giant phospholipid vesicles (ATP production rate up to ∼100 ATP∙s−1 per ATP synthase). The chromatophore morphology and the orientation of the photophosphorylation proteins were characterized by cryo-electron microscopy (cryo-EM) and time-resolved spectroscopy. The freshly synthesized ATP has been employed for sustaining the transcription of a DNA gene, following the RNA biosynthesis inside individual vesicles by confocal microscopy. The hybrid multicompartment approach here proposed is very promising for the construction of full-fledged artificial protocells because it relies on easy-to-obtain and ready-to-use chromatophores, paving the way for artificial simplified-autotroph protocells (ASAPs).

Author(s):  
Emiliano Altamura ◽  
Paola Albanese ◽  
Roberto Marotta ◽  
Francesco Milano ◽  
Michele Fiore ◽  
...  

AbstractThe construction of energetically autonomous artificial protocells is one of the most urgent and challenging requirements in bottom-up synthetic biology. Here we show a hybrid multi-compartment approach to build Artificial Simplified-Autotroph Protocells (ASAPs) in an effective manner. Chromatophores obtained from Rhodobacter sphaeroides accomplish the photophosphorylation of ADP to ATP functioning as nanosized photosynthetic organellae when encapsulated inside artificial giant phospholipid vesicles. Under continuous illumination chromatophores produce ATP that in turn sustains the transcription of a DNA gene by T7 RNA polymerase inside ASAPs. Cryo-EM and time-resolved spectroscopy were used for characterizing the chromatophore morphology and the orientation of the photophosphorylation proteins, which allow high ATP production rates (up to ~100 ATP/s per ATP synthase). mRNA biosynthesis inside individual vesicles has been determined by confocal microscopy. The hybrid multi-compartment approach here proposed appears at the same time convenient and effective, and thus very promising for the construction of full-fledged artificial protocells.


2009 ◽  
Vol 96 (3) ◽  
pp. 411a-412a
Author(s):  
Zonghuan Lu ◽  
David Barnard ◽  
Tanvir R. Shaikh ◽  
Hisham Mohamed ◽  
Xing Meng ◽  
...  

2009 ◽  
Vol 168 (3) ◽  
pp. 388-395 ◽  
Author(s):  
Zonghuan Lu ◽  
Tanvir R. Shaikh ◽  
David Barnard ◽  
Xing Meng ◽  
Hisham Mohamed ◽  
...  

1991 ◽  
Vol 161 (1) ◽  
pp. 47-57 ◽  
Author(s):  
Jean Lepault ◽  
Inge Erk ◽  
Gisèle Nicolas ◽  
Jean-Luc Ranck

Science ◽  
2017 ◽  
Vol 358 (6365) ◽  
pp. 936-940 ◽  
Author(s):  
Hui Guo ◽  
Stephanie A. Bueler ◽  
John L. Rubinstein

Mitochondrial adenosine triphosphate (ATP) synthase produces the majority of ATP in eukaryotic cells, and its dimerization is necessary to create the inner membrane folds, or cristae, characteristic of mitochondria. Proton translocation through the membrane-embedded FO region turns the rotor that drives ATP synthesis in the soluble F1 region. Although crystal structures of the F1 region have illustrated how this rotation leads to ATP synthesis, understanding how proton translocation produces the rotation has been impeded by the lack of an experimental atomic model for the FO region. Using cryo–electron microscopy, we determined the structure of the dimeric FO complex from Saccharomyces cerevisiae at a resolution of 3.6 angstroms. The structure clarifies how the protons travel through the complex, how the complex dimerizes, and how the dimers bend the membrane to produce cristae.


2008 ◽  
Vol 14 (S2) ◽  
pp. 1598-1599
Author(s):  
Z Lu ◽  
J McMahon ◽  
H Mohamed ◽  
D Barnard ◽  
TR Shaikh ◽  
...  

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008


2014 ◽  
Vol 24 (11) ◽  
pp. 115001 ◽  
Author(s):  
Zonghuan Lu ◽  
David Barnard ◽  
Tanvir R Shaikh ◽  
Xing Meng ◽  
Carmen A Mannella ◽  
...  

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