title: Dimethyl sulphoxide and Ca2+ stimulate assembly of Vibrio cholerae FtsZ.
abstract: 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 Ca(2+) 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.
authors: Department of Biotechnology and Dr. B.C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta, 35 Ballygunge Circular Road Kolkata, WB 700019, India.
authors: Abhisek
authors: A
authors: Chatterjee
authors: Department of Biotechnology and Dr. B.C. Guha Centre for Genetic Engineering and Biotechnology, University of Calcutta, 35 Ballygunge Circular Road Kolkata, WB 700019, India. Electronic address: gcbcg@caluniv.ac.in.
authors: Gopal
authors: G
authors: Chakrabarti
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002771:Cholera / Q000188:drug therapy / Q000235:genetics* / Q000382:microbiology; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D004926:Escherichia coli; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006801:Humans; D055550:Protein Stability; D014734:Vibrio cholerae / Q000737:chemistry / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D020558:GTP Phosphohydrolases; D002118:Calcium; D004121:Dimethyl Sulfoxide
keywords: Calcium ion and DMSO; Cloning; Expression and purification; FtsZ; Polymerization; Vibrio cholerae