Product uses
- Identification and characterization of FtsZ binding proteins
- Characterization of FtsZ dynamics
- Developing FtsZ ligands that may be used as anti-bacterial agents.
MaterialFtsZ is a bacterial cytoskeletal protein that is essential for cell division many prokaryotes (1). It has been shown to be a bacterial homolog of eukaryotic tubulin, based both on a low sequence identity and a striking structural similarity (2). Just like eukaryotic tubulin, FtsZ polymerizes as well as binds and hydrolyzes GTP in a polymerization dependent manner.
Streptococcus pneumoniae FtsZ protein has been purified after over-expression in E.coli. S.p. FtsZ has a 6xHis tag and an approximate molecular weight of 45 kDa (Fig. 1). FTZ03 is provided as a lyophilized protein with buffer and stabilizers.
PurityProtein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 10% polyacrylamide gel. FtsZ protein was calculated to be >90% pure (see Figure 1, Lane 2).
PurityPurity is determined by scanning densitometry of proteins on SDS-PAGE gels. Samples are >90% pure. No free dye is apparent in the final product.
Figure 1: A 100 µg sample of S.p. FtsZ protein was separated on a 4-20% gradient SDS-PAGE gel and stained with Coomassie blue. Lane 1, 100µg FTZ03. SeeBlue molecular weight markers are from Invitrogen.
Biological activityThe biological activity of FtsZ can be determined in two ways, first from its ability to efficiently polymerize into protofilaments and sheets in vitro in the presence of Mg2+ and GTP, and secondly to hydrolyze GTP to GDP and Pi. Polymerization with Mg2+:GTP in the presence of a crowding reagent (e.g. carbohydrate polymers Ficoll or dextran in "Sheet Buffer") results in sheets of FtsZ protofilaments that sediment at 100,000xg, analysis of the pellet and sup by protein assay or SDS-PAGE indicates the proportion of polymerizable FtsZ protein; the batch of protein passes Quality Control when the pelleted amount is greater than 60% of the total and the supernatnat protein is in agreement with its known critical concentration for assembly. GTPase activity is determined in a "Protofilament Buffer" by measuring the Pi released over time by using the MESEG phosphate detection kit (Cat# BK052) or an endpoint assay (Cat. # BK054). The batch of protein passes Quality Control when a specific activity greater than 0.05 mol GTP / mol FtsZ / min is obtained in Protofilament Buffer.
For more information about FtsZ proteins, Click here.
References
- Bramhill, D. (1997) Bacterial cell division. Annu. Rev. Cell Dev. Biol. 13, 395-424.
- Erickson, H. P. (1998) Atomic structures of tubulin and FtsZ. Trends Cell Biol. 8, 133-137.
For product Datasheets and MSDSs please click on the PDF links below. For additional information, click on the FAQs tab above or contact our Technical Support department at tservice@cytoskeleton.com
Coming soon! For the most recent publications citing this product, please contact our Technical Service department at tservice@cytoskeleton.com
Question 1: Given the high sequence homology between FtsZ proteins, is it necessary to use a species-specific FtsZ matter?
Answer 1: FtsZ sequence homology between some species is high e.g. S.enterica and E.coli, while most others have 40-70% homology. Even with high homology, if the divergent sequences encode a binding domain for FtsZ accessory proteins or novel compounds/drugs, binding of the proteins and/or test compounds could vary significantly. Whether using FtsZ as a drug discovery tool for novel antibiotic research or for better understanding bacterial cell division, it is best to use FtsZ proteins that are specific for the bacterial species you are studying to insure reliable and consistent results.
Question 2: What is the proper way to store the FtsZ protein (Cat. # FTZ03)?
Answer 2: The lyophilized protein should be stored at 4°C with fresh desiccant where it will be stable for 6 months. Storage at -70°C is also an option, though the rubber seal on the container can crack at this temperature and allow in moisture. Therefore we recommend storing at 4°C. If stored at -70°C, it is imperative to include desiccant with the lyophilized protein if this storage condition is utilized. After reconstituting as directed, the concentrated protein should then be aliquoted and snap frozen in liquid nitrogen. Store these aliquots at -70°C for 6 months. For working concentrations, further dilution of the protein should be made with MES Buffer (100 mM MES pH6.5, 1mM EGTA). NOTE: It is very important to snap freeze the FtsZ protein in liquid nitrogen as other methods of freezing will result in significantly reduced activity. Defrost rapidly by placing in a room temperature water bath for 1 min. Avoid repeated freeze/thaw cycles.
If you have any questions concerning this product, please contact our Technical Service department at tservice@cytoskeleton.com
Cytoskeleton公司成立于1993年，专注于生物化学和细胞过程研究中的纯化蛋白和便捷试剂盒开发与生产。公司提供药物筛选、信号转导、细胞骨架研究相关的系列试剂盒和产品，尤其以细胞骨架相关研究见长，既能满足于样品较少的科学研究，也可以用于小规模筛选研究和高通量大规模筛选研究。此外，公司还提供微管蛋白，肌动蛋白，小G蛋白，GAPs，GEFs等现有产品的药物筛选服务。
本网站将在规定时间内给予删除等相关处理。
