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2016 ; 55
(9
): 1301-13
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Contributions of Coulombic and Hofmeister Effects to the Osmotic Activation of
Escherichia coli Transporter ProP
#MMPMID26871755
Culham DE
; Shkel IA
; Record MT Jr
; Wood JM
Biochemistry
2016[Mar]; 55
(9
): 1301-13
PMID26871755
show ga
Osmosensing transporters mediate osmolyte accumulation to forestall cellular
dehydration as the extracellular osmolality increases. ProP is a bacterial
osmolyte-H(+) symporter, a major facilitator superfamily member, and a paradigm
for osmosensing. ProP activity is a sigmoid function of the osmolality. It is
determined by the osmolality, not the magnitude or direction of the osmotic
shift, in cells and salt-loaded proteoliposomes. The activation threshold varies
directly with the proportion of anionic phospholipid in cells and
proteoliposomes. The osmosensory mechanism was probed by varying the salt
composition and concentration outside and inside proteoliposomes. Data analysis
was based on the hypothesis that the fraction of maximal transporter activity at
a particular luminal salt concentration reflects the proportion of ProP molecules
in an active conformation. ProP attained the same activity at the same osmolality
when diverse, membrane-impermeant salts were added to the external medium.
Contributions of Coulombic and/or Hofmeister salt effects to ProP activation were
examined by varying the luminal salt cation (K(+) and Na(+)) and anion (chloride,
phosphate, and sulfate) composition and then systematically increasing the
luminal salt concentration by increasing the external osmolality. ProP activity
increased with the sixth power of the univalent cation concentration, independent
of the type of anion. This indicates that salt activation of ProP is a Coulombic,
cation effect resulting from salt cation accumulation and not site-specific
cation binding. Possible origins of this Coulombic effect include folding or
assembly of anionic cytoplasmic ProP domains, an increase in local membrane
surface charge density, and/or the juxtaposition of anionic protein and membrane
surfaces during activation.