Analysis of Protein Dynamics with Tandem Fluorescent Protein Timers

2014 ◽  
pp. 195-210 ◽  
Author(s):  
Anton Khmelinskii ◽  
Michael Knop
Keyword(s):  
Protein Dynamics ◽  
Fluorescent Protein

scholarly journals Imaging Synaptic Protein Dynamics Using Photoactivatable Green Fluorescent Protein

2012 ◽  
Vol 2012 (7) ◽  
pp. pdb.prot070029-pdb.prot070029
Author(s):  
R. M. Weimer ◽  
T. C. Hill ◽  
A. M. Hamilton ◽  
K. Zito
Keyword(s):  
Green Fluorescent Protein ◽  
Protein Dynamics ◽  
Fluorescent Protein ◽  
Synaptic Protein ◽  
Green Fluorescent

scholarly journals Monitoring Protein Dynamics in Protein O-Mannosyltransferase Mutants In Vivo by Tandem Fluorescent Protein Timers

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2622 ◽  
Author(s):  
Joan Castells-Ballester ◽  
Ewa Zatorska ◽  
Matthias Meurer ◽  
Patrick Neubert ◽  
Anke Metschies ◽  
...  
Keyword(s):  
Protein Dynamics ◽  
Secretory Pathway ◽  
Fluorescent Protein ◽  
Model Organism ◽  
Protein Glycosylation ◽  
Major Protein ◽  
Specific Substrate ◽  
Evolutionarily Conserved ◽  
The Impact

For proteins entering the secretory pathway, a major factor contributing to maturation and homeostasis is glycosylation. One relevant type of protein glycosylation is O-mannosylation, which is essential and evolutionarily-conserved in fungi, animals, and humans. Our recent proteome-wide study in the eukaryotic model organism Saccharomyces cerevisiae revealed that more than 26% of all proteins entering the secretory pathway receive O-mannosyl glycans. In a first attempt to understand the impact of O-mannosylation on these proteins, we took advantage of a tandem fluorescent timer (tFT) reporter to monitor different aspects of protein dynamics. We analyzed tFT-reporter fusions of 137 unique O-mannosylated proteins, mainly of the secretory pathway and the plasma membrane, in mutants lacking the major protein O-mannosyltransferases Pmt1, Pmt2, or Pmt4. In these three pmtΔ mutants, a total of 39 individual proteins were clearly affected, and Pmt-specific substrate proteins could be identified. We observed that O-mannosylation may cause both enhanced and diminished protein abundance and/or stability when compromised, and verified our findings on the examples of Axl2-tFT and Kre6-tFT fusion proteins. The identified target proteins are a valuable resource towards unraveling the multiple functions of O-mannosylation at the molecular level.

Photoactivated structural dynamics of fluorescent proteins

2012 ◽  
Vol 40 (3) ◽  
pp. 531-538 ◽  
Author(s):  
Dominique Bourgeois ◽  
Aline Regis-Faro ◽  
Virgile Adam
Keyword(s):  
Protein Dynamics ◽  
Structural Dynamics ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Super Resolution ◽  
Simulation Tools ◽  
X Ray Crystallography ◽  
Hybrid Approaches ◽  
Actinic Light ◽  
Green Fluorescent

Proteins of the GFP (green fluorescent protein) family have revolutionized life sciences because they allow the tagging of biological samples in a non-invasive genetically encoded way. ‘Phototransformable’ fluorescent proteins, in particular, have recently attracted widespread interest, as their fluorescence state can be finely tuned by actinic light, a property central to the development of super-resolution microscopy. Beyond microscopy applications, phototransformable fluorescent proteins are also exquisite tools to investigate fundamental protein dynamics. Using light to trigger processes such as photoactivation, photoconversion, photoswitching, blinking and photobleaching allows the exploration of the conformational landscape in multiple directions. In the present paper, we review how structural dynamics of phototransformable fluorescent proteins can be monitored by combining X-ray crystallography, in crystallo optical spectroscopy and simulation tools such as quantum chemistry/molecular mechanics hybrid approaches. Besides their usefulness to rationally engineer better performing fluorescent proteins for nanoscopy and other biotechnological applications, these investigations provide fundamental insights into protein dynamics.

scholarly journals Protein dynamics control proton transfer from bulk solvent to protein interior: A case study with a green fluorescent protein

2005 ◽  
Vol 14 (7) ◽  
pp. 1787-1799 ◽  
Author(s):  
Anoop M. Saxena ◽  
Jayant B. Udgaonkar ◽  
Guruswamy Krishnamoorthy
Keyword(s):  
Green Fluorescent Protein ◽  
Proton Transfer ◽  
Protein Dynamics ◽  
Fluorescent Protein ◽  
Green Fluorescent

scholarly journals Fluorescent protein tools for studying protein dynamics in living cells: a review

2008 ◽  
Vol 13 (3) ◽  
pp. 031202 ◽  
Author(s):  
Richard N. Day ◽  
Fred Schaufele
Keyword(s):  
Protein Dynamics ◽  
Fluorescent Protein ◽  
Living Cells

Tandem fluorescent protein timers for in vivo analysis of protein dynamics

2012 ◽  
Vol 30 (7) ◽  
pp. 708-714 ◽  
Author(s):  
Anton Khmelinskii ◽  
Philipp J Keller ◽  
Anna Bartosik ◽  
Matthias Meurer ◽  
Joseph D Barry ◽  
...  
Keyword(s):  
Protein Dynamics ◽  
Fluorescent Protein ◽  
In Vivo Analysis
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