pppGpp is a highly phosphorylated guanosine, which has an additional phosphate residue at the ribose 5' position. pppGpp is a global regulatory nucleotide. It binds to several protein targets to regulate multiple biochemical events in bacterial physiology from core central principles to various metabolic pathways. pppGpp has a linear phosphate chain that provides additional flexibility to accommodate the various conformations according to stereo binding sites of the respective target proteins to bind proteins of various functional classes. Initially, pppGpp was found to be a transcriptional regulatory molecule because it binds to RNA polymerase and regulates transcriptional gene regulation. In the past decade, several other target proteins of pppGpp have been identified. To date, more than 30 proteins have been reported to be regulated by the binding of signal nucleotides. Regulation of different biochemical activities by pppGpp requires fine molecular interactions with different classes of proteins so that different functions can be regulated.
Fig.1 The structure and ball stick model of pppGpp. (Kushwaha, et al., 2020)
pppGpp has attracted wide attention because of its unique biological functions. CD BioGlyco provides a variety of different services to synthesize pppGpp in vivo and in vitro. High purity of the products we synthesize, at the gram or milligram level. The synthesis methods provided by our company are as follows:
In a cell-free synthesis system, synthesized by pppGpp synthetase transfers the pyrophosphate group from ATP to GDP/GTP to form. There are two main types of pppGpp synthetases, multidomain long RelA type, and small alarm synthases. Ribosomes can be recovered, stored, and repeatedly used as a source of synthetic activity. For the synthesized products, chromatography, Ultraviolet (UV) scanning, and X-ray testing are performed to provide customers with all the product test reports.
CD BioGlyco uses GDP, GTP, and ATP as reaction substrates and E. coli as raw material for product synthesis and adds protein inhibitors in the synthesis process to achieve the purpose of delaying bacterial contamination. The reaction process was monitored by visualization of the substrate and the product, and the nucleotide product of the reaction was determined during the subsequent purification process. For product purification separation, we eluted using the double primary method and removed it thoroughly. The intermolecular interactions between pppGpp and target proteins were characterized by various physicochemical parameters, including hydrogen bonds, van der Waals interactions, aromatic stacking, and side chains of protein-interacting residues.
Fig.2 The synthesis pathway of pppGpp. (CD BioGlyco)
CD BioGlyco has many years of successful experience in the field of Carbohydrate Synthesis. Moreover, we provide efficient cAMP, cGMP, FAD, FMN, NAD, and NADP production services. Our company has always been well received by clients with quality service and reliable products. If you are interested in our service, please feel free to contact us further to introduce the specific service in detail.
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