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Biophys J. 2014 May 6;106(9):L33-5. doi: 10.1016/j.bpj.2014.03.019.

Tracking single serotonin transporter molecules at the endoplasmic reticulum and plasma membrane.

Author information

1
Institute of Applied Physics, Vienna University of Technology, Vienna, Austria.
2
Institute of Applied Physics, Vienna University of Technology, Vienna, Austria. Electronic address: enricoklotzsch@gmail.com.
3
European Molecular Biology Laboratory, Heidelberg, Germany.
4
Center for Physiology and Pharmacology, Institute of Pharmacology, Medical University Vienna, Vienna, Austria.
5
Institute of Applied Physics, Vienna University of Technology, Vienna, Austria. Electronic address: schuetz@iap.tuwien.ac.at.

Abstract

Transmembrane proteins are synthesized and folded in the endoplasmic reticulum (ER), an interconnected network of flattened sacs or tubes. Up to now, this organelle has eluded a detailed analysis of the dynamics of its constituents, mainly due to the complex three-dimensional morphology within the cellular cytosol, which precluded high-resolution, single-molecule microscopy approaches. Recent evidences, however, pointed out that there are multiple interaction sites between ER and the plasma membrane, rendering total internal reflection microscopy of plasma membrane proximal ER regions feasible. Here we used single-molecule fluorescence microscopy to study the diffusion of the human serotonin transporter at the ER and the plasma membrane. We exploited the single-molecule trajectories to map out the structure of the ER close to the plasma membrane at subdiffractive resolution. Furthermore, our study provides a comparative picture of the diffusional behavior in both environments. Under unperturbed conditions, the majority of proteins showed similar mobility in the two compartments; at the ER, however, we found an additional 15% fraction of molecules moving with 25-fold faster mobility. Upon degradation of the actin skeleton, the diffusional behavior in the plasma membrane was strongly influenced, whereas it remained unchanged in the ER.

PMID:
24806941
PMCID:
PMC4017317
DOI:
10.1016/j.bpj.2014.03.019
[Indexed for MEDLINE]
Free PMC Article
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