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Strain Engineering for Phosphoprotein

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CD Biosynsis is committed to combining genomics and proteomics to produce high-quality phosphoprotein products to better assist in the study of the mechanisms between protein phosphorylation and molecular regulation.

Background of Synthetic Biology-Driven Phosphoprotein Production

Protein phosphorylation occurs as one of the most efficient regulatory pathways in biology for the vast majority of proteins. As a result of the addition of phosphate groups to the amino acid side chains of proteins, the conformation, activity, and properties of proteins interacting with other molecules are altered. Protein phosphorylation is involved in the regulation of cell signaling, gene expression, cell division, and many other biological processes. Exploring the mystery of phosphorylation modifications has become a key concern for biologists. The production of phosphorylated proteins using synthetic biology techniques relies mainly on exogenous protein expression systems. However, phosphorylated proteins produced by eukaryotic expression systems are difficult to isolate and purify, and common prokaryotic expression systems produce target proteins without post-translational modifications. Therefore, the search for novel biosynthetic methods and improved protein expression systems is of great importance for the synthesis of phosphoproteins.

Figure 1. Accurate protein synthesis and conventional methodology to alter aaRS specificity. (Vargas-Rodriguez O, et al., 2018)Figure 1. Accurate protein synthesis and conventional methodology to alter aaRS specificity. (Vargas-Rodriguez O, et al., 2018)

What We Provide

CD Biosynsis is dedicated to the production of different types of phosphoproteins based on our synthetic biology platform using strategies such as phosphate amino acid residue mimics, cell-free protein synthesis, and genetic codon expansion.

Featured Services

  • We are able to synthesize phosphoproteins using solid-phase peptide synthesis techniques and peptide fragment ligation techniques.
  • We are able to use natural chemical linkages to ligate synthetic phosphoproteins.
  • We are able to incorporate phosphorylated amino acid residues or their mimics into the protein using a genetic code extension strategy.

Deliverables

  • Phosphoproteins.
  • Phosphorylated amino acid residue mimics.

How We Can Help

Semi-Synthesis of Phosphoproteins

We can produce a recombinant protein containing a C-terminal thioester by a protein expression system, which is then linked to a peptide containing phosphorylated residues by a chemical ligation reaction. This semi-synthetic approach allows for the construction of larger proteins and the production of recombinant proteins at a lower cost. We have used semi-synthetic methods such as expression protein ligation to produce a variety of phosphoproteins.

Synthesis of Phosphorylated Amino Acid Residue Mimics

Phosphoproteins cannot be biosynthesized without phosphorylated amino acid residues. Our scientists have worked to develop a series of mimics that can replace natural phosphorylated amino acid residues for the synthesis of phosphoproteins. Phosphorylation of proteins occurs mainly at amino acid residues such as serine, threonine, and tyrosine. So far, we have successfully synthesized the following phosphoproteins using phosphorylated amino acid residue mimics.

Phosphorylated amino acid residue mimic Phosphoprotein
pSer Mitogen-activated protein kinase kinases 1
With no lysine kinase-4
Phosphorylated green fluorescent protein
pThr Cell cycle protein-dependent kinase 2
pTyr Signal transducer and activator of transcription 1
Myoglobin

Post-Modification of Proteins

We are able to phosphorylate specific amino acid residues of the recombinant protein by kinase-catalyzed reaction activation chemistry.

Cell-Free Phosphoprotein Synthesis

We are able to use cell-free protein synthesis techniques to produce phosphoproteins in cell lysate species using phosphorylated amino acid residues or their mimics.

Applications of Phosphoprotein

CD Biosynsis can develop tailored tools and customized approaches to harness the power of synthetic biology to drive phosphoprotein production and meet the needs of customers in a variety of industries.

  • As a probe for protein phosphorylation networks.
  • As biosensor for detecting structural changes in proteins.
  • For the development of cancer therapeutic agents.
  • For the study of neurodegenerative diseases.

Want to Learn More?

CD Biosynsis provides the most comprehensive and efficient solutions for synthetic biology workflows. We are committed to helping our customers solve all problems encountered in phosphoprotein production to advance their applications in a wide range of fields. Each of our deliverables will undergo a rigorous quality inspection test to ensure the reliability and accuracy of the results. If you are interested in our services or have any further questions, please do not hesitate to contact us.

References

  1. Duan HZ, et al. Unremitting progresses for phosphoprotein synthesis. Curr Opin Chem Biol. 2020 Oct; 58: 96-111.
  2. Chen Z, Cole PA. Synthetic approaches to protein phosphorylation. Curr Opin Chem Biol. 2015 Oct; 28: 115-22.
  3. Vargas-Rodriguez O, et al. Upgrading aminoacyl-tRNA synthetases for genetic code expansion. Curr Opin Chem Biol. 2018 Oct; 46: 115-122.

Please note that all services are for research use only. Not intended for any clinical use.

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