Locating the Band III protein in quasi-native cell membranes

Band III is a key protein for the structure and function of red blood cell membranes. To date, the distribution and morphology of Band III in cell membranes is still unclear because of limited approaches. We applied Topography and RECognition imaging microscopy (TREC), which extends the application of atomic force microscopy (AFM) to recognize a single molecule in a biological complex, to visually locate a single Band III protein in quasi-native cell membranes by anti-Band III-functionalized AFM tips under physiological conditions. The Band III proteins are well distributed in the inner leaflet of cell membranes. The height of the whole Band III protein in cell membranes is in the range of 9–13 nm. The unbinding force between Band III in the membrane and anti-Band III on the AFM tip is about 70 pN with the loading rate at 40 nN/s. Our result is significant in revealing the location and morphology of Band III in the inner cell membrane at the molecular level.

[1]  D. Lohr,et al.  Single-molecule recognition imaging microscopy. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[2]  W. Kühlbrandt,et al.  Three‐dimensional map of the dimeric membrane domain of the human erythrocyte anion exchanger, Band 3. , 1994, The EMBO journal.

[3]  D. Jay.  Role of Band 3 in Homeostasis and Cell Shape , 1996, Cell.

[4]  J. Gimsa,et al.  Do band 3 protein conformational changes mediate shape changes of human erythrocytes? , 1995, Molecular membrane biology.

[5]  Hao Yan,et al.  Recognition imaging with a DNA aptamer. , 2006, Biophysical journal.

[6]  Y. Dufrêne,et al.  Detection and localization of single molecular recognition events using atomic force microscopy , 2006, Nature Methods.

[7]  Nikolai Gaponik,et al.  THIOL-CAPPING OF CDTE NANOCRYSTALS: AN ALTERNATIVE TO ORGANOMETALLIC SYNTHETIC ROUTES , 2002 .

[8]  L. Cantley,et al.  Structural aspects of the red cell anion exchange protein. , 1986, Annual review of biochemistry.

[9]  D. Branton,et al.  Visualization of the protein associations in the erythrocyte membrane skeleton. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[10]  N. Mohandas,et al.  Red cell membrane: past, present, and future. , 2008, Blood.

[11]  T. Lange,et al.  Determination of CFTR densities in erythrocyte plasma membranes using recognition imaging , 2008, Nanotechnology.

[12]  R. Harrington,et al.  Kinked DNA , 1997, Nature.

[13]  W. Kühlbrandt,et al.  Two‐dimensional structure of the membrane domain of human band 3, the anion transport protein of the erythrocyte membrane. , 1993, The EMBO journal.

[14]  P. Hinterdorfer,et al.  Localization of the ergtoxin-1 receptors on the voltage sensing domain of hERG K+ channel by AFM recognition imaging , 2008, Pflügers Archiv - European Journal of Physiology.

[15]  Gerald Kada,et al.  Properties of lipid microdomains in a muscle cell membrane visualized by single molecule microscopy , 2000, The EMBO journal.

[16]  M. Tanner,et al.  Structure of the anion-transport protein of the human erythrocyte membrane. Further studies on the fragments produced by proteolytic digestion. , 1979, The Biochemical journal.

[17]  P. Low,et al.  Crystallographic structure and functional interpretation of the cytoplasmic domain of erythrocyte membrane band 3. , 2000, Blood.

[18]  Andreas Ebner,et al.  Reduced number of CFTR molecules in erythrocyte plasma membrane of cystic fibrosis patients , 2006, Molecular membrane biology.

[19]  L. K. Drickamer.  Fragmentation of the 95,000-dalton transmembrane polypeptide in human erythrocyte membranes. , 1976, The Journal of biological chemistry.

[20]  M. Mann,et al.  Deep Coverage Mouse Red Blood Cell Proteome , 2008, Molecular & Cellular Proteomics.

[21]  D. Lohr,et al.  Two-component atomic force microscopy recognition imaging of complex samples. , 2007, Analytical biochemistry.

[22]  H. Schillers.  Imaging CFTR in its native environment , 2008, Pflügers Archiv - European Journal of Physiology.

[23]  Xin Shang,et al.  Localization of Na+-K+ ATPases in quasi-native cell membranes. , 2009, Nano letters.

[24]  Y. Lyubchenko,et al.  Atomic force microscopy of long DNA: imaging in air and under water. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. Tanner,et al.  Functional cell surface expression of the anion transport domain of human red cell band 3 (AE1) in the yeast Saccharomyces cerevisiae. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[26]  D. Branton,et al.  The molecular basis of erythrocyte shape. , 1986, Science.