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Journal of Cell Science, Vol 113, Issue 22 3931-3938, Copyright © 2000 by Company of Biologists
JOURNAL ARTICLES |
U Rescher, N Zobiack and V Gerke
Center for Molecular Biology of Inflammation, Institute for Medical Biochemistry, von-Esmarchstrasse 56, Germany.
Annexin 1 is a Ca(2+)-regulated membrane binding protein and a major substrate of the epidermal growth factor receptor kinase. Because of its properties and intracellular distribution, the protein has been implicated in endocytic trafficking of the receptor, in particular in receptor sorting occurring in multivesicular endosomes. Up to now, however, the localization of annexin 1 to cellular membranes has been limited to subcellular fractionation and immunocytochemical analyses of fixed cells. To establish its localization in live cells, we followed the intracellular fate of annexin 1 molecules fused to the Green Fluorescent Protein (GFP). We show that annexin 1-GFP associates with distinct, transferrin receptor-positive membrane structures in living HeLa cells. A GFP chimera containing the Ca(2+)/phospholipid-binding protein core of annexin 1 also shows a punctate intracellular distribution, although the structures labeled here do not resemble early but, at least in part, late endosomes. In contrast, the cores of annexins 2 and 4 fused to GFP exhibit a cytoplasmic or a different punctate distribution, respectively, indicating that the highly homologous annexin core domains carry distinct membrane specificities within live cells. By inactivating the three high-affinity Ca(2+) binding sites in annexin 1 we also show that endosomal membrane binding of the protein in live HeLa cells depends on the integrity of these Ca(2+) binding sites. More detailed analysis identifies a single Ca(2+) site in the second annexin repeat that is crucially involved in establishing the membrane association. These results reveal for the first time that intracellular membrane binding of an annexin in living cells requires Ca(2+) and is mediated in part through an annexin core domain that is capable of establishing specific interactions.
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  A. K. McNeil, U. Rescher, V. Gerke, and P. L. McNeil Requirement for Annexin A1 in Plasma Membrane Repair J. Biol. Chem., November 17, 2006; 281(46): 35202 - 35207. [Abstract] [Full Text] [PDF]
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 M. Eisenhut and U. Haberkorn Molecular Position of Radiolabels and Its Impact on Functional Integrity of Proteins J. Nucl. Med., September 1, 2006; 47(9): 1400 - 1402. [Full Text] [PDF]
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 A. Piljic and C. Schultz Annexin A4 Self-Association Modulates General Membrane Protein Mobility in Living Cells Mol. Biol. Cell, July 1, 2006; 17(7): 3318 - 3328. [Abstract] [Full Text] [PDF]
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 U. Rescher, D. Ruhe, C. Ludwig, N. Zobiack, and V. Gerke Annexin 2 is a phosphatidylinositol (4,5)-bisphosphate binding protein recruited to actin assembly sites at cellular membranes J. Cell Sci., July 15, 2004; 117(16): 3473 - 3480. [Abstract] [Full Text] [PDF]
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 J. C. Mills, N. Andersson, C. V. Hong, T. S. Stappenbeck, and J. I. Gordon Molecular characterization of mouse gastric epithelial progenitor cells PNAS, November 12, 2002; 99(23): 14819 - 14824. [Abstract] [Full Text] [PDF]
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 V. Gerke and S. E. Moss Annexins: From Structure to Function Physiol Rev, April 1, 2002; 82(2): 331 - 371. [Abstract] [Full Text] [PDF]
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N. Zobiack, U. Rescher, S. Laarmann, S. Michgehl, M. A. Schmidt, and V. Gerke Cell-surface attachment of pedestal-forming enteropathogenic E. coli induces a clustering of raft components and a recruitment of annexin 2 J. Cell Sci., January 1, 2002; 115(1): 91 - 98. [Abstract] [Full Text] [PDF]
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