Dimethyl sulphoxide and Ca2+ stimulate assembly of Vibrio cholerae FtsZ

作者：Chatterjee Abhisek; Chakrabarti Gopal^*

来源：Biochimie, 2014, 105: 64-75.

DOI：10.1016/j.biochi.2014.06.013

• FtsZ
• Vibrio cholerae
• Cloning
• Expression and purification
• Polymerization
• Calcium ion and DMSO

We cloned, overexpressed and purified Vibrio cholerae FtsZ protein for the first time. We used several complementary techniques to probe and compare the comparative assembly properties of recombinant Vibrio cholerae FtsZ (VcFtsZ) and Escherichia coli FtsZ (EcFtsZ). We observed that VcFtsZ polymerized at a slower rate than EcFtsZ and interestingly its polymerization was highly dependent on the presence of Ca2+ ion. Furthermore, DMSO specifically modulated the polymerization of VcFtsZ, promoted polymer bundling and increased the stability of the VcFtsZ protofilaments. Whereas DMSO showed no significant stimulatory effect on the assembly and bundling of EcFtsZ. Transmission electron microscopy experiments demonstrated that in presence of 8% DMSO the average thickness of the VcFtsZ polymers were increased significantly. DMSO specifically stabilized the VcFtsZ polymers against dilution induced disassembly and it reduced the GTPase activity of VcFtsZ. These results collectively suggested that despite lot of sequence homology, the assembly of VcFtsZ and EcFtsZ are differently regulated processes. We expect to use this knowledge of assembly properties of VcFtsZ for screening of small molecules against VcFtsZ for development of anti-cholera agent.

• 出版日期2014-10