Plasma membrane protein clusters appear in CFTR-expressing Xenopus laevis oocytes after cAMP stimulation

The Journal of Membrane Biology
Hermann SchillersH Oberleithner

Abstract

Membrane trafficking of the cystic fibrosis transmembrane conductance regulator (CFTR) is supposed to be an important mechanism controlled by the intracellular messenger cAMP. This has been shown with fluorescence techniques, electron microscopy and membrane capacitance measurements. In order to visualize protein insertion we applied atomic force microscopy (AFM) to inside-out oriented plasma membrane patches of CFTR-expressing Xenopus laevis oocytes before and after cAMP-stimulation. In a first step, oocytes injected with CFTR-cRNA were voltage-clamped, verifying successful CFTR expression. Water-injected oocytes served as controls. Then, plasma membrane patches were excised, placed (inside out) on glass and scanned by AFM. Before cAMP-stimulation plasma membranes of both water-injected and CFTR-expressing oocytes contained about 200 proteins per micron 2. Molecular protein masses were estimated from molecular volumes measured by AFM. Before cAMP-stimulation, protein distribution showed a peak value of 11 nm protein height corresponding to 475 kDa. During cAMP-stimulation with 1 mM isobutylmethylxanthine (IBMX) plasma membrane protein density increased in water-injected oocytes to 700 proteins per micron 2 while the peak value...Continue Reading

Citations

Dec 7, 2007·Pflügers Archiv : European journal of physiology·Hermann Schillers
Jun 9, 2006·Journal of the Association for Research in Otolaryngology : JARO·Michio MurakoshiHiroshi Wada
Aug 13, 2008·The Journal of Biological Chemistry·Christopher M LewarchikRaymond A Frizzell
Feb 16, 2005·Annual Review of Physiology·John R Riordan
Dec 15, 2006·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·Katrin KlicheHans Oberleithner
Sep 1, 2007·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·Christina H LimBen C Tilly
Aug 23, 2006·Molecular Membrane Biology·Tobias LangeHermann Schillers
Apr 12, 2002·FEBS Letters·Andrei SegalWolf Michael Weber
Dec 31, 2002·Journal of Receptor and Signal Transduction Research·Michael HortonPetri Lehenkari
Aug 19, 2004·Journal of Cellular Physiology·Victor ShahinHans Oberleithner
May 2, 2003·British Journal of Pharmacology·Alan W Cuthbert
Jul 29, 2006·Journal of Microscopy·M SantacroceG Poletti
Dec 15, 2006·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·Astrid StumpfHermann Schillers
Aug 4, 2006·Microscopy Research and Technique·F OrsiniG Poletti
Jan 4, 2008·Microscopy Research and Technique·Massimo SantacroceGiulio Poletti
Oct 21, 2005·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Thomas Ludwig

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