Deuterium oxide promotes assembly and bundling of FtsZ protofilaments.
The assembly and bundling of FtsZ protofilaments play an important role during bacterial cell division. Deuterium oxide (D2O) is known to have strong stabilization effects on the assembly dynamics of several proteins including tubulin, a homologue of FtsZ. Here, we found that D2O enhanced the light-scattering intensity of the assembly reaction, increased sedimentable polymer mass, and induced bundling of FtsZ protofilaments. D2O also increased the stability of FtsZ polymers under challenged GTP conditions and suppressed dilution-induced disassembly of protofilaments. D2O enhances the assembly parameters of FtsZ and microtubules albeit differently. For example, D2O induced bundling of FtsZ protofilaments, whereas it did not induce bundling of microtubules in vitro. In addition, D2O strongly suppressed the GTP hydrolysis rate of microtubules, but it had no effect on the initial rate of GTP hydrolysis of the FtsZ assembly. D2O (80%) also increased the helical content of FtsZ by 25% compared to the helical content of FtsZ in aqueous buffer. D2O was shown to reduce the binding of 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (bis-ANS) to tubulin. In contrast, we found that D2O strongly enhanced the binding of bis-ANS to FtsZ. The results indicated that D2O promotes assembly and bundling of FtsZ protofilaments by increasing hydrophobic interactions between the protofilaments. The results also suggest that the phosphate release rather than the on-site GTP hydrolysis is the rate-limiting step of the GTP turnover reaction.
Santra
Manas Kumar
MK
School of Biosciences and Bioengineering, Indian Institute of Technology, Bombay, Powai, Mumbai, India.
Dasgupta
Debjani
D
Panda
Dulal
D
D002942:Circular Dichroism; D017666:Deuterium Oxide / Q000494:pharmacology*; D004795:Enzyme Stability; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008027:Light; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008958:Models, Molecular; D012542:Scattering, Radiation
D029968:Escherichia coli Proteins; C076835:FtsZ84 protein, E coli; D046911:Macromolecular Substances; D017666:Deuterium Oxide