PMID- 27821772
OWN - NLM
STAT- MEDLINE
DCOM- 20180416
LR  - 20181113
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 113
IP  - 47
DP  - 2016 Nov 22
TI  - Emulating proton-induced conformational changes in the vesicular monoamine 
      transporter VMAT2 by mutagenesis.
PG  - E7390-E7398
AB  - Neurotransporters located in synaptic vesicles are essential for communication 
      between nerve cells in a process mediated by neurotransmitters. Vesicular 
      monoamine transporter (VMAT), a member of the largest superfamily of 
      transporters, mediates transport of monoamines to synaptic vesicles and storage 
      organelles in a process that involves exchange of two H(+) per substrate. VMAT 
      transport is inhibited by the competitive inhibitor reserpine, a second-line 
      agent to treat hypertension, and by the noncompetitive inhibitor tetrabenazine, 
      presently in use for symptomatic treatment of hyperkinetic disorders. During the 
      transport cycle, VMAT is expected to occupy at least three different 
      conformations: cytoplasm-facing, occluded, and lumen-facing. The lumen- to 
      cytoplasm-facing transition, facilitated by protonation of at least one of the 
      essential membrane-embedded carboxyls, generates a binding site for reserpine. 
      Here we have identified residues in the cytoplasmic gate and show that mutations 
      that disrupt the interactions in this gate also shift the equilibrium toward the 
      cytoplasm-facing conformation, emulating the effect of protonation. These 
      experiments provide significant insight into the role of proton translocation in 
      the conformational dynamics of a mammalian H(+)-coupled antiporter, and also 
      identify key aspects of the mode of action and binding of two potent inhibitors 
      of VMAT2: reserpine binds the cytoplasm-facing conformation, and tetrabenazine 
      binds the lumen-facing conformation.
FAU - Yaffe, Dana
AU  - Yaffe D
AD  - Department of Biological Chemistry, Alexander Silberman Institute of Life 
      Sciences, Hebrew University, Jerusalem 91904, Israel.
FAU - Vergara-Jaque, Ariela
AU  - Vergara-Jaque A
AD  - Computational Structural Biology Section, National Institute of Neurological 
      Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.
FAU - Forrest, Lucy R
AU  - Forrest LR
AUID- ORCID: 0000-0003-1855-7985
AD  - Computational Structural Biology Section, National Institute of Neurological 
      Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.
FAU - Schuldiner, Shimon
AU  - Schuldiner S
AUID- ORCID: 0000-0002-4874-6237
AD  - Department of Biological Chemistry, Alexander Silberman Institute of Life 
      Sciences, Hebrew University, Jerusalem 91904, Israel; 
      shimon.schuldiner@huji.ac.il.
LA  - eng
GR  - R01 NS016708/NS/NINDS NIH HHS/United States
GR  - R56 NS016708/NS/NINDS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, N.I.H., Intramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20161107
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Protons)
RN  - 0 (Slc18a2 protein, rat)
RN  - 0 (Vesicular Monoamine Transport Proteins)
RN  - 8B1QWR724A (Reserpine)
RN  - Z9O08YRN8O (Tetrabenazine)
SB  - IM
MH  - Animals
MH  - Binding Sites
MH  - Cytoplasm/genetics/metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Models, Molecular
MH  - *Mutation
MH  - Protein Conformation
MH  - Protons
MH  - Rats
MH  - Reserpine/*metabolism
MH  - Tetrabenazine/*metabolism
MH  - Vesicular Monoamine Transport Proteins/*chemistry/*genetics/metabolism
PMC - PMC5127352
OTO - NOTNLM
OT  - ion coupling
OT  - neurotransmitter transporter
OT  - reserpine
OT  - tetrabenazine
COIS- The authors declare no conflict of interest.
EDAT- 2016/11/09 06:00
MHDA- 2018/04/17 06:00
PMCR- 2017/05/22
CRDT- 2016/11/09 06:00
PHST- 2016/11/09 06:00 [pubmed]
PHST- 2018/04/17 06:00 [medline]
PHST- 2016/11/09 06:00 [entrez]
PHST- 2017/05/22 00:00 [pmc-release]
AID - 1605162113 [pii]
AID - 201605162 [pii]
AID - 10.1073/pnas.1605162113 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2016 Nov 22;113(47):E7390-E7398. doi: 
      10.1073/pnas.1605162113. Epub 2016 Nov 7.