NDLI: Phosphatidylinositol 4,5-bisphosphate (PIP2) stimulates the electrogenic Na/HCO3 cotransporter NBCe1-A expressed in Xenopus oocytes

Content Provider	PubMed Central
Author	Wu, Jianping Mcnicholas, Carmel M. Bevensee, Mark O.
Abstract	Bicarbonate transporters are regulated by signaling molecules/ions such as protein kinases, ATP, and Ca2+. While phospholipids such as PIP2 can stimulate Na-H exchanger activity, little is known about phospholipid regulation of bicarbonate transporters. We used the patch-clamp technique to study the function and regulation of heterologously expressed rat NBCe1-A in excised macropatches from Xenopus laevis oocytes. Exposing the cytosolic side of inside-out macropatches to a 5% CO2/33 mM HCO3 − solution elicited a mean inward current of 14 pA in 74% of macropatches attached to pipettes (−Vp = −60 mV) containing a low-Na+, nominally HCO3 −-free solution. The current was 80–90% smaller in the absence of Na+, approximately 75% smaller in the presence of 200 μM DIDS, and absent in macropatches from H2O-injected oocytes. NBCe1-A currents exhibited time-dependent rundown that was inhibited by removing Mg2+ in the presence or absence of vanadate and F− to reduce general phosphatase activity. Applying 5 or 10 μM PIP2 (diC8) in the presence of HCO3 − induced an inward current in 54% of macropatches from NBC-expressing, but not H2O-injected oocytes. PIP2-induced currents were HCO3 −-dependent and somewhat larger following more NBCe1-A rundown, 62% smaller in the absence of Na+, and 90% smaller in the presence of 200 μM DIDS. The polycation neomycin (250–500 μM) reduced the PIP2-induced inward current by 69%; spermine (100 μM) reduced the current by 97%. Spermine, poly-D-lysine, and neomycin all reduced the baseline HCO3 −-induced inward currents by as much as 85%. In summary, PIP2 stimulates NBCe1-A activity, and phosphoinositides are regulators of bicarbonate transporters.
Related Links	http://dx.doi.org/10.1073/pnas.0906303106
Starting Page	14150
File Format	PDF
ISSN	10916490
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	33
Volume Number	106
Language	English
Publisher	National Academy of Sciences
Publisher Date	2009-08-18
Access Restriction	Open
Rights Holder	National Academy of Sciences
Subject Keyword	Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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