scholarly journals Correction: Live cell single molecule tracking and localization microscopy of bioorthogonally labeled plasma membrane proteins

Nanoscale ◽  
2020 ◽  
Vol 12 (35) ◽  
pp. 18476-18477
Author(s):  
Andres I. König ◽  
Raya Sorkin ◽  
Ariel Alon ◽  
Dikla Nachmias ◽  
Kalyan Dhara ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Single Molecule ◽  
Live Cell ◽  
Plasma Membrane Proteins ◽  
Single Molecule Tracking ◽  
Localization Microscopy

Correction for ‘Live cell single molecule tracking and localization microscopy of bioorthogonally labeled plasma membrane proteins’ by Andres I. König et al., Nanoscale, 2020, 12, 3236–3248, DOI: 10.1039/C9NR08594G.

scholarly journals Live cell single molecule tracking and localization microscopy of bioorthogonally labeled plasma membrane proteins

Nanoscale ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 3236-3248 ◽  
Author(s):  
Andres I. König ◽  
Raya Sorkin ◽  
Ariel Alon ◽  
Dikla Nachmias ◽  
Kalyan Dhara ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Genetic Code ◽  
Single Molecule ◽  
Live Cell ◽  
Bioorthogonal Chemistry ◽  
Plasma Membrane Proteins ◽  
Single Molecule Tracking ◽  
Localization Microscopy ◽  
Genetic Code Expansion

Visualizing the spatiotemporal organization of plasma membrane proteins at the nanoscale by combining live-cell single-molecule applications with direct fluorescent dye labeling via genetic code expansion and bioorthogonal chemistry.

scholarly journals Live cell single molecule tracking and localization microscopy of bioorthogonally labeled plasma membrane proteins

2019 ◽  
Author(s):  
Andres I. König ◽  
Raya Sorkin ◽  
Ariel Alon ◽  
Dikla Nachmias ◽  
Kalyan Dhara ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Single Molecule ◽  
Egf Receptor ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Protein Labeling ◽  
Bioorthogonal Chemistry ◽  
Plasma Membrane Proteins ◽  
Localization Microscopy

ABSRACTTracking the localization and mobility of individual proteins in live cells is key for understanding how they mediate their function. Such information can be obtained from single molecule imaging techniques such as Single Particle Tracking (SPT) and Single Molecule Localization Microscopy (SMLM). Genetic code expansion (GCE) combined with bioorthogonal chemistry offers an elegant approach for direct labeling of proteins with fluorescent dyes, holding great potential for improving protein labeling in single molecule applications. Here we calibrated conditions for performing SPT and live-SMLM of bioorthogonally labeled plasma membrane proteins in live mammalian cells. Using SPT, the diffusion of bioorthogonally labeled EGF receptor and the prototypical Shaker voltage-activated potassium channel (Kv) was measured and characterized. Applying live-SMLM to bioorthogonally labeled Shaker Kv channels enabled visualizing the plasma membrane distribution of the channel over time with ~30 nm accuracy. Finally, by competitive labeling with two Fl-dyes, SPT and live-SMLM were performed in a single cell and both the density and dynamics of the EGF receptor were measured at single molecule resolution in subregions of the cell. We conclude that GCE and bioorthogonal chemistry is a highly suitable, flexible approach for protein labeling in quantitative single molecule applications that outperforms current protein live-cell labeling approaches.

Detergent fractionation with subsequent subtractive suppression hybridization as a tool for identifying genes coding for plasma membrane proteins

2009 ◽  
Vol 18 (6) ◽  
pp. 527-535 ◽  
Author(s):  
Andreas Lange ◽  
Claudia Kistler ◽  
Tanja B. Jutzi ◽  
Alexandr V. Bazhin ◽  
Claus Detlev Klemke ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Plasma Membrane Proteins ◽  
Subtractive Suppression Hybridization

scholarly journals pH-dependent Cargo Sorting from the Golgi

2011 ◽  
Vol 286 (12) ◽  
pp. 10058-10065 ◽  
Author(s):  
Chunjuan Huang ◽  
Amy Chang
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Atpase Activity ◽  
Secretory Pathway ◽  
Ph Dependence ◽  
Glucose Deprivation ◽  
Ph Homeostasis ◽  
Plasma Membrane Proteins ◽  
Cytosolic Ph ◽  
Cargo Sorting

The vacuolar proton-translocating ATPase (V-ATPase) plays a major role in organelle acidification and works together with other ion transporters to maintain pH homeostasis in eukaryotic cells. We analyzed a requirement for V-ATPase activity in protein trafficking in the yeast secretory pathway. Deficiency of V-ATPase activity caused by subunit deletion or glucose deprivation results in missorting of newly synthesized plasma membrane proteins Pma1 and Can1 directly from the Golgi to the vacuole. Vacuolar mislocalization of Pma1 is dependent on Gga adaptors although no Pma1 ubiquitination was detected. Proper cell surface targeting of Pma1 was rescued in V-ATPase-deficient cells by increasing the pH of the medium, suggesting that missorting is the result of aberrant cytosolic pH. In addition to mislocalization of the plasma membrane proteins, Golgi membrane proteins Kex2 and Vrg4 are also missorted to the vacuole upon loss of V-ATPase activity. Because the missorted cargos have distinct trafficking routes, we suggest a pH dependence for multiple cargo sorting events at the Golgi.

scholarly journals 5-Iodonaphthyl-1-azide labeling of plasma membrane proteins adjacent to specific sites via energy transfer

1997 ◽  
Vol 1324 (2) ◽  
pp. 320-332 ◽  
Author(s):  
Bruce I Meiklejohn ◽  
Noorulhuda A Rahman ◽  
Deborah A Roess ◽  
B.George Barisas
Keyword(s):  
Plasma Membrane ◽  
Energy Transfer ◽  
Membrane Proteins ◽  
Plasma Membrane Proteins
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