PMID- 31393114
OWN - NLM
STAT- MEDLINE
DCOM- 20200715
LR  - 20200715
IS  - 2379-3694 (Electronic)
IS  - 2379-3694 (Linking)
VI  - 4
IP  - 9
DP  - 2019 Sep 27
TI  - Genetically Encoded Halide Sensor-Based Fluorescent Assay for Rapid Screening of 
      Glutamate Transport and Inhibition.
PG  - 2358-2366
LID - 10.1021/acssensors.9b00944 [doi]
AB  - Glutamate is the main excitatory neurotransmitter in the mammalian central 
      nervous system. Excitatory amino acid transporters (EAATs) are a family of 
      transmembrane transporters responsible for glutamate uptake into cells, and their 
      malfunction is related to a variety of diseases, including neurodegenerative 
      diseases and stroke. Screening for and developing inhibitors of EAATs as well as 
      related transporters is a significant field of study for biomedical and 
      pharmaceutical applications. Rapid, high-throughput methods are critical for the 
      study of glutamate transporters, and fluorescent methods are appealing for this 
      purpose as compared to more traditional electrophysiological methods. In this 
      study, we present a method for studying glutamate transporters and inhibitors by 
      utilizing a mutated version of a yellow fluorescent protein (YFP) highly 
      sensitive to quenching by anions (mClY). We applied this YFP variant to 
      fluorescent imaging of anion flux in HEK293 cells caused by transiently expressed 
      excitatory amino acid carrier 1 (EAAC1) and excitatory amino acid transporter 2 
      (EAAT2) and its inhibition by competitive blockers. This method enables rapid 
      identification of inhibitors and, potentially, activators of EAAT function, which 
      is critical for glutamate transport research.
FAU - Zielewicz, Laura
AU  - Zielewicz L
AD  - Department of Chemistry , Binghamton University , 4400 Vestal Parkway East , 
      Binghamton , New York 13902 , United States.
FAU - Grewer, Christof
AU  - Grewer C
AUID- ORCID: 0000-0002-8342-9878
AD  - Department of Chemistry , Binghamton University , 4400 Vestal Parkway East , 
      Binghamton , New York 13902 , United States.
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20190821
PL  - United States
TA  - ACS Sens
JT  - ACS sensors
JID - 101669031
RN  - 0 (Bacterial Proteins)
RN  - 0 (Excitatory Amino Acid Transporter 2)
RN  - 0 (Excitatory Amino Acid Transporter 3)
RN  - 0 (Halogens)
RN  - 0 (Luminescent Proteins)
RN  - 0 (SLC1A1 protein, human)
RN  - 0 (yellow fluorescent protein, Bacteria)
RN  - 3KX376GY7L (Glutamic Acid)
SB  - IM
MH  - Bacterial Proteins/genetics
MH  - Biological Transport
MH  - Excitatory Amino Acid Transporter 2/antagonists & inhibitors/genetics/metabolism
MH  - Excitatory Amino Acid Transporter 3/antagonists & inhibitors/genetics/metabolism
MH  - Glutamic Acid/*metabolism
MH  - HEK293 Cells
MH  - Halogens/*metabolism
MH  - Humans
MH  - Luminescent Proteins/genetics
MH  - Optical Imaging/*methods
MH  - Time Factors
OTO - NOTNLM
OT  - binding
OT  - electrophysiology
OT  - fluorescence
OT  - glutamate transporter
OT  - inhibitor
OT  - method development
EDAT- 2019/08/09 06:00
MHDA- 2020/07/16 06:00
CRDT- 2019/08/09 06:00
PHST- 2019/08/09 06:00 [pubmed]
PHST- 2020/07/16 06:00 [medline]
PHST- 2019/08/09 06:00 [entrez]
AID - 10.1021/acssensors.9b00944 [doi]
PST - ppublish
SO  - ACS Sens. 2019 Sep 27;4(9):2358-2366. doi: 10.1021/acssensors.9b00944. Epub 2019 
      Aug 21.