Creative BioMart is a well-known expert who is committed to incorporating a variety of non-canonical amino acids into proteins to introduce new functions, redesign natural proteins and artificial protein macromolecules. With years of experience, we provide customized services for engineering photocrosslinked proteins to precisely meet customer requirements.

Introduction of Photoreactive Amino Acids

Unnatural amino acids with photoreactive groups, such as benzophenones, arylazides, and naphthalenes. Photoactivation of these species with UV light produces a reactive carbene intermediate that irreversibly cross-links proteins with zero-length bond distances on any amino acid side chain or peptide backbone. Introducing cross-linkable side chains into proteins provides a complementary approach to elucidating specific interactions in vivo. In recent years, remarkable achievements have been made in the research of photoreactive amino acids. For example photocaged cysteine, serine and tyrosine residues have been introduced site-specifically to study proteins involved in different cellular processes. Unnatural and photoreactive amino acids as biochemical probes for immune function.

Fig 1. Application of proteins carrying unnatural amino acids containing novel functional group. (Hohsaka T, et al., 2002)

Services

The main advantage of photocrosslinking is the higher crosslinking specificity due to the shorter lifetime of the excited intermediates. It has been widely used to covalently fix interactions in living cells. As a leading service provider of protein engineering, Creative BioMart has successfully synthesized two unique photo-reactive amino acid derivatives: Photo-Leucine and Photo-Methionine, which are structurally similar to their respective natural amino acids, allowing for their performance in living cells protein cross-linking.

Protein-protein interactions are key to organizing cellular processes in space and time. The only direct way to identify this interaction in their cellular context is through photocrosslinking. We provide one-stop service for photocrosslinking proteins:

• Photocrosslinking service using photoreactive amino acids: we use UV light to irreversibly photocrosslink proteins within protein interaction domains containing photoreactive amino acids (methionine, leucine, cysteine, serine, and tyrosine).
• In vivo crosslinking service for live cells: we treated HeLa cells with photocrosslinking proteins prior to photoactivation with UV light and analyzed by SDS-PAGE and Western blotting using an antibody against the homodimeric transcription factor Stat3.

Creative BioMart is committed to providing global customers with unique protein-specific crosslinkers for living cells, helping you capture more diverse protein-protein interactions and better recover proteins after cell lysis. We will work with you to develop the most appropriate strategy and provide the most meaningful data for your research for accelerating the research of life sciences. If you are interested in our services, please do not hesitate to contact us for more information.

References

1. Suchanek M, Radzikowska A, Thiele C. (2005) Photo-leucine and photo-methionine allow identification of protein-protein interactions in living cells[J]. Nature methods. 2(4): 261-268.
2. Hohsaka T, Sisido M. (2002) Incorporation of non-natural amino acids into proteins[J]. Current opinion in chemical biology. 6(6): 809-815.

• Protein Engineering Based on Backbone Amides
• Protein Engineering Based on Fluorescent Amino Acids
• Protein Engineering Based on Fluorinated Amino Acids