PMID: 9178623Jul 1, 1997Paper

Imaging excised apical plasma membrane patches of MDCK cells in physiological conditions with atomic force microscopy

Pflügers Archiv : European journal of physiology
J LärmerHans Oberleithner

Abstract

We combined the patch-clamp technique with atomic force microscopy (AFM) to visualize plasma membrane proteins protruding from the extracellular surface of cultured kidney cells (MDCK cells). To achieve molecular resolution, patches were mechanically isolated from whole MDCK cells by applying the patch-clamp technique. The excised inside-out patches were transferred on freshly cleaved mica and imaged with the AFM in air and under physiological conditions (i. e. in fluid). Thus, the resolution could be increased considerably (lateral and vertical resolutions 5 and 0.1 nm, respectively) as compared to experiments on intact cells, where plasma membrane proteins were hardly detectable. The apical plasma membrane surface of the MDCK cells showed multiple protrusions which could be identified as membrane proteins through the use of pronase. These proteins had a density of about 90 per micron(2), with heights between 1 and 9 nm, and lateral dimensions of 20-60 nm. Their frequency distribution showed a peak value of 3 nm for the protein height. A simplified assumption - modelling plasma membrane proteins as spherical structures protruding from the lipid bilayer - allowed an estimation of the possible molecular weights of these proteins...Continue Reading

Citations

May 22, 2009·European Biophysics Journal : EBJ·Francesco OrsiniF V Sacchi
Dec 7, 2007·Pflügers Archiv : European journal of physiology·Hermann Schillers
Aug 5, 2008·Pflügers Archiv : European journal of physiology·Michio MurakoshiHiroshi Wada
Oct 8, 2009·Pflügers Archiv : European journal of physiology·Ghanshyam P SinhaKuni H Iwasa
Jun 9, 2006·Journal of the Association for Research in Otolaryngology : JARO·Michio MurakoshiHiroshi Wada
Oct 22, 1998·FEBS Letters·U ZieglerP Groscurth
Apr 18, 2002·British Journal of Pharmacology·Calum S NeishJ Michael Edwardson
Aug 4, 2006·Microscopy Research and Technique·F OrsiniG Poletti
Mar 4, 2005·Journal of Cellular Physiology·Stephan KusickThomas Ludwig
Feb 27, 2009·Small·Bärbel LorenzAndreas Janshoff
Jan 20, 2005·The Journal of Biological Chemistry·Nelson P BarreraJ Michael Edwardson
May 12, 2000·American Journal of Physiology. Renal Physiology·R M Henderson, H Oberleithner
Jul 24, 1998·Biophysical Journal·C Le GrimellecJ P Goudonnet

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