PMID- 25546576
OWN - NLM
STAT- MEDLINE
DCOM- 20160205
LR  - 20181113
IS  - 1097-4547 (Electronic)
IS  - 0360-4012 (Print)
IS  - 0360-4012 (Linking)
VI  - 93
IP  - 7
DP  - 2015 Jul
TI  - Displacing hexokinase from mitochondrial voltage-dependent anion channel impairs 
      GLT-1-mediated glutamate uptake but does not disrupt interactions between GLT-1 
      and mitochondrial proteins.
PG  - 999-1008
LID - 10.1002/jnr.23533 [doi]
AB  - The glutamate transporter GLT-1 is the major route for the clearance of 
      extracellular glutamate in the forebrain, and most GLT-1 protein is found in 
      astrocytes. This protein is coupled to the Na(+) electrochemical gradient, 
      supporting the active intracellular accumulation of glutamate. We recently used a 
      proteomic approach to identify proteins that may interact with GLT-1 in rat 
      cortex, including the Na(+)/K(+) -ATPase, most glycolytic enzymes, and several 
      mitochondrial proteins. We also showed that most GLT-1 puncta ( approximately  70%) are 
      overlapped by mitochondria in astroglial processes in organotypic slices. From 
      this analysis, we proposed that the glycolytic enzyme hexokinase (HK)-1 might 
      physically form a scaffold to link GLT-1 and mitochondria because HK1 is known to 
      interact with the outer mitochondrial membrane protein voltage-dependent anion 
      channel (VDAC). The current study validates the interactions among HK-1, VDAC, 
      and GLT-1 by using forward and reverse immunoprecipitations and provides evidence 
      that a subfraction of HK1 colocalizes with GLT-1 in vivo. A peptide known to 
      disrupt the interaction between HK and VDAC did not disrupt interactions between 
      GLT-1 and several mitochondrial proteins. In parallel experiments, displacement 
      of HK from VDAC reduced GLT-1-mediated glutamate uptake. These results suggest 
      that, although HK1 forms coimmunoprecipitatable complexes with both VDAC and 
      GLT-1, it does not physically link GLT-1 to mitochondrial proteins. However, the 
      interaction of HK1 with VDAC supports GLT-1-mediated transport activity.
CI  - (c) 2014 Wiley Periodicals, Inc.
FAU - Jackson, Joshua G
AU  - Jackson JG
AD  - Children's Hospital of Philadelphia Research Institute, Philadelphia, 
      Pennsylvania.
AD  - Department of Pediatrics, University of Pennsylvania, Philadelphia, Pennsylvania.
FAU - O'Donnell, John C
AU  - O'Donnell JC
AD  - Children's Hospital of Philadelphia Research Institute, Philadelphia, 
      Pennsylvania.
AD  - Department of Pharmacology, University of Pennsylvania, Philadelphia, 
      Pennsylvania.
FAU - Krizman, Elizabeth
AU  - Krizman E
AD  - Children's Hospital of Philadelphia Research Institute, Philadelphia, 
      Pennsylvania.
FAU - Robinson, Michael B
AU  - Robinson MB
AD  - Children's Hospital of Philadelphia Research Institute, Philadelphia, 
      Pennsylvania.
AD  - Department of Pediatrics, University of Pennsylvania, Philadelphia, Pennsylvania.
AD  - Department of Pharmacology, University of Pennsylvania, Philadelphia, 
      Pennsylvania.
LA  - eng
GR  - P30 HD026979/HD/NICHD NIH HHS/United States
GR  - T32 GM008076/GM/NIGMS NIH HHS/United States
GR  - F31 NS086255/NS/NINDS NIH HHS/United States
GR  - T32 NS007413/NS/NINDS NIH HHS/United States
GR  - R01NS077773/NS/NINDS NIH HHS/United States
GR  - R01 NS077773/NS/NINDS NIH HHS/United States
GR  - P30 HD26979/HD/NICHD NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20141226
PL  - United States
TA  - J Neurosci Res
JT  - Journal of neuroscience research
JID - 7600111
RN  - 0 (Excitatory Amino Acid Transporter 2)
RN  - 0 (Mitochondrial Proteins)
RN  - 0 (Slc1a2 protein, rat)
RN  - 0 (Voltage-Dependent Anion Channels)
RN  - 3KX376GY7L (Glutamic Acid)
RN  - EC 2.7.1.1 (Hexokinase)
SB  - IM
MH  - Animals
MH  - Cerebral Cortex/*metabolism/ultrastructure
MH  - Excitatory Amino Acid Transporter 2/*metabolism
MH  - Glutamic Acid/*metabolism
MH  - Hexokinase/chemistry/*metabolism
MH  - Immunoprecipitation
MH  - Male
MH  - Mitochondrial Membranes/metabolism
MH  - Mitochondrial Proteins/*metabolism
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Synaptosomes/metabolism
MH  - Voltage-Dependent Anion Channels/*metabolism
PMC - PMC4441544
MID - NIHMS646828
OTO - NOTNLM
OT  - glutamate transporter
OT  - hexokinase
OT  - mitochondria
EDAT- 2014/12/30 06:00
MHDA- 2016/02/06 06:00
PMCR- 2016/07/01
CRDT- 2014/12/30 06:00
PHST- 2014/09/16 00:00 [received]
PHST- 2014/11/11 00:00 [revised]
PHST- 2014/11/30 00:00 [accepted]
PHST- 2014/12/30 06:00 [entrez]
PHST- 2014/12/30 06:00 [pubmed]
PHST- 2016/02/06 06:00 [medline]
PHST- 2016/07/01 00:00 [pmc-release]
AID - 10.1002/jnr.23533 [doi]
PST - ppublish
SO  - J Neurosci Res. 2015 Jul;93(7):999-1008. doi: 10.1002/jnr.23533. Epub 2014 Dec 
      26.