Inhibition of assembly of bacterial cell division protein FtsZ by the hydrophobic dye 5,5'-bis-(8-anilino-1-naphthalenesulfonate).
To gain further insight into the structural relatedness of tubulin and FtsZ, the tubulin-like prokaryotic cell division protein, we tested the effect of tubulin assembly inhibitors on FtsZ assembly. Common tubulin inhibitors, such as colchicine, colcemid, benomyl, and vinblastine, had no effect on Ca2+-promoted GTP-dependent assembly of FtsZ into polymers. However, the hydrophobic probe 5, 5'-bis-(8-anilino-1-naphthalenesulfonate) (bis-ANS) inhibited FtsZ assembly. The potential mechanisms for inhibition are discussed. Titrations of FtsZ with bis-ANS indicated that FtsZ has one high affinity binding site and multiple low affinity binding sites. ANS (8-anilino-1-naphthalenesulfonate), a hydrophobic probe similar to bis-ANS, had no inhibitory effect on FtsZ assembly. Because tubulin assembly has also been shown to be inhibited by bis-ANS but not by ANS, it supports the idea that FtsZ and tubulin share similar conformational properties. Ca2+, which promotes GTP-dependent FtsZ assembly, stimulated binding of bis-ANS or ANS to FtsZ, suggesting that Ca2+ binding induces changes in the hydrophobic conformation of the protein. Interestingly, depletion of bound Ca2+ with EGTA further enhanced bis-ANS fluorescence. These findings suggest that both binding and dissociation of Ca2+ are capable of inducing FtsZ conformational changes, and these changes could promote the GTP-dependent assembly of FtsZ.
Yu
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Department of Microbiology and Molecular Genetics, University of Texas Medical School, Houston, Texas 77030, USA.
Margolin
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D000817:Anilino Naphthalenesulfonates / Q000378:metabolism / Q000494:pharmacology*; D001419:Bacteria / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D001704:Biopolymers; D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects*; D003598:Cytoskeletal Proteins; D005456:Fluorescent Dyes / Q000378:metabolism / Q000494:pharmacology*; D006160:Guanosine Triphosphate / Q000378:metabolism; D015335:Molecular Probes; D011485:Protein Binding; D011487:Protein Conformation
D000817:Anilino Naphthalenesulfonates; D001426:Bacterial Proteins; D001704:Biopolymers; D003598:Cytoskeletal Proteins; D005456:Fluorescent Dyes; C071525:FtsZ protein, Bacteria; D015335:Molecular Probes; C027132:1-anilino-8-naphthalenesulfonate; D006160:Guanosine Triphosphate; D002118:Calcium