PMID- 25336661
OWN - NLM
STAT- MEDLINE
DCOM- 20150303
LR  - 20240322
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 289
IP  - 49
DP  - 2014 Dec 5
TI  - Functionally important carboxyls in a bacterial homologue of the vesicular 
      monoamine transporter (VMAT).
PG  - 34229-40
LID - 10.1074/jbc.M114.607366 [doi]
AB  - Transporters essential for neurotransmission in mammalian organisms and bacterial 
      multidrug transporters involved in antibiotic resistance are evolutionarily 
      related. To understand in more detail the evolutionary aspects of the 
      transformation of a bacterial multidrug transporter to a mammalian 
      neurotransporter and to learn about mechanisms in a milieu amenable for 
      structural and biochemical studies, we identified, cloned, and partially 
      characterized bacterial homologues of the rat vesicular monoamine transporter 
      (rVMAT2). We performed preliminary biochemical characterization of one of them, 
      Brevibacillus brevis monoamine transporter (BbMAT), from the bacterium B. brevis. 
      BbMAT shares substrates with rVMAT2 and transports them in exchange with >1H(+), 
      like the mammalian transporter. Here we present a homology model of BbMAT that 
      has the standard major facilitator superfamily fold; that is, with two domains of 
      six transmembrane helices each, related by 2-fold pseudosymmetry whose axis runs 
      normal to the membrane and between the two halves. The model predicts that four 
      carboxyl residues, a histidine, and an arginine are located in the transmembrane 
      segments. We show here that two of the carboxyls are conserved, equivalent to the 
      corresponding ones in rVMAT2, and are essential for H(+)-coupled transport. We 
      conclude that BbMAT provides an excellent experimental paradigm for the study of 
      its mammalian counterparts and bacterial multidrug transporters.
CI  - (c) 2014 by The American Society for Biochemistry and Molecular Biology, Inc.
FAU - Yaffe, Dana
AU  - Yaffe D
AD  - From the Department of Biological Chemistry, Alexander Silberman Institute of 
      Life Sciences, Hebrew University, 91904 Jerusalem, Israel.
FAU - Vergara-Jaque, Ariela
AU  - Vergara-Jaque A
AD  - the Computational Structural Biology Section, NINDS, National Institutes of 
      Health, Bethesda, Maryland 20852, and.
FAU - Shuster, Yonatan
AU  - Shuster Y
AD  - From the Department of Biological Chemistry, Alexander Silberman Institute of 
      Life Sciences, Hebrew University, 91904 Jerusalem, Israel.
FAU - Listov, Dina
AU  - Listov D
AD  - From the Department of Biological Chemistry, Alexander Silberman Institute of 
      Life Sciences, Hebrew University, 91904 Jerusalem, Israel.
FAU - Meena, Sitaram
AU  - Meena S
AD  - the Department of Cellular and Molecular Physiology, Yale University School of 
      Medicine, New Haven, Connecticut 06520.
FAU - Singh, Satinder K
AU  - Singh SK
AD  - the Department of Cellular and Molecular Physiology, Yale University School of 
      Medicine, New Haven, Connecticut 06520.
FAU - Forrest, Lucy R
AU  - Forrest LR
AD  - the Computational Structural Biology Section, NINDS, National Institutes of 
      Health, Bethesda, Maryland 20852, and.
FAU - Schuldiner, Shimon
AU  - Schuldiner S
AD  - From the Department of Biological Chemistry, Alexander Silberman Institute of 
      Life Sciences, Hebrew University, 91904 Jerusalem, Israel, 
      shimon.schuldiner@huji.ac.il.
LA  - eng
SI  - PDB/3WDO
GR  - R21 MH098180/MH/NIMH NIH HHS/United States
GR  - R21MH098180/MH/NIMH NIH HHS/United States
GR  - NS16708/NS/NINDS NIH HHS/United States
GR  - R56 NS016708/NS/NINDS NIH HHS/United States
GR  - R01 NS016708/NS/NINDS NIH HHS/United States
GR  - R00 MH083050/MH/NIMH NIH HHS/United States
GR  - R01 MH100688/MH/NIMH NIH HHS/United States
GR  - ImNIH/Intramural 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
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20141021
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Bacterial Proteins)
RN  - 0 (Biogenic Monoamines)
RN  - 0 (Carrier Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Slc18a2 protein, rat)
RN  - 0 (Vesicular Monoamine Transport Proteins)
RN  - 4QD397987E (Histidine)
RN  - 94ZLA3W45F (Arginine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Arginine/chemistry/metabolism
MH  - Bacterial Proteins/*chemistry/genetics/metabolism
MH  - Biogenic Monoamines/*chemistry/metabolism
MH  - Brevibacillus/*chemistry/genetics/metabolism
MH  - Carrier Proteins/*chemistry/genetics/metabolism
MH  - Drug Resistance, Bacterial
MH  - Escherichia coli/genetics/metabolism
MH  - Evolution, Molecular
MH  - Gene Expression
MH  - Histidine/chemistry/metabolism
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Protein Folding
MH  - Rats
MH  - Recombinant Proteins/chemistry/genetics/metabolism
MH  - Sequence Alignment
MH  - Structural Homology, Protein
MH  - Structure-Activity Relationship
MH  - Substrate Specificity
MH  - Synaptic Transmission/physiology
MH  - Vesicular Monoamine Transport Proteins/*chemistry/genetics/metabolism
PMC - PMC4256354
OTO - NOTNLM
OT  - Antibiotic Resistance
OT  - Membrane Transport
OT  - Multidrug Transporter
OT  - Neurotransmitter
OT  - Neurotransmitter Transport
OT  - Proton Transport
EDAT- 2014/10/23 06:00
MHDA- 2015/03/04 06:00
PMCR- 2015/12/05
CRDT- 2014/10/23 06:00
PHST- 2014/10/23 06:00 [entrez]
PHST- 2014/10/23 06:00 [pubmed]
PHST- 2015/03/04 06:00 [medline]
PHST- 2015/12/05 00:00 [pmc-release]
AID - S0021-9258(20)47370-3 [pii]
AID - M114.607366 [pii]
AID - 10.1074/jbc.M114.607366 [doi]
PST - ppublish
SO  - J Biol Chem. 2014 Dec 5;289(49):34229-40. doi: 10.1074/jbc.M114.607366. Epub 2014 
      Oct 21.