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Microbiology (Reading) ; 142 ( Pt 5): 1133-1140, 1996 May.
Article in English | MEDLINE | ID: mdl-8704955

ABSTRACT

Actin has been described in all eukaryotic cells as the major microfilament cytoskeletal protein. Although prokaryotic cells do not have a cytoskeleton, proteins related to the latter have been found in different prokaryotic species. We have found prokaryotic actin-related proteins in the enterobacterium Escherichia coli and in the cyanobacteria Anabaena cylindrica and Anabaena variabilis. They were identified by the following criteria: (1) by cross-reaction with a fluorescent conjugated anti-actin (rat-brain) mAb by Western blot analysis (in total cellular extracts); (2) specific binding of acetone powder and soluble cellular extracts to DNase I; and (3) specific binding of cells and total cellular extracts to phalloidin. In E coli, specific binding of phalloidin labelled with rhodamine to cells was detected by spectrofluorometry. In total cellular extracts, three bands of 60, 43 and 35 kDa were weakly recognized by the mAb by Western blot analysis; this recognition increased when phalloidin was added to the extracts. Furthermore, three polypeptides of kDa were isolated by binding to DNase I, showing pI values of 6.7, 6.65 and 6.6, less acidic than all reported actin pI values. In A. cylindrica and A. variabilis, specific binding of phalloidin labelled with rhodamine to cells was also detected by spectrofluorometry. In total and soluble cellular extracts, the mAb recognized two bands of 45 and 40 kDa by Western blot analysis, but only the first was purified by binding to DNase I, and it showed three isoforms of pI values 6.8, 6.5 and 6.4. These results suggest the presence, in prokaryotes, of proteins with similar biochemical characteristics to eukaryotic actin.


Subject(s)
Actins/chemistry , Anabaena/chemistry , Bacterial Proteins/isolation & purification , Escherichia coli/chemistry , Plant Proteins/isolation & purification , Actins/immunology , Anabaena/immunology , Anabaena/ultrastructure , Animals , Antibodies, Bacterial/immunology , Bacterial Proteins/chemistry , Bacterial Proteins/immunology , Biopolymers , Blotting, Western , Cross Reactions , Deoxyribonuclease I/metabolism , Escherichia coli/immunology , Escherichia coli/ultrastructure , Evolution, Molecular , Phalloidine/metabolism , Plant Proteins/chemistry , Plant Proteins/immunology , Protein Binding , Rats , Species Specificity
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