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Apr 1, 2008 (Vol. 28, No. 7)

Studying Protein Interactions in Living Cells

Method for Protein Crosslinking Utilizes Photoreactive Amino Acids

  • Protein solubility in cell lysates after photoreactive amino acid crosslinking resulted in 95% protein recovery compared to only 35% and 65% protein recovery after treatment with formaldehyde and NHS-ester crosslinkers, respectively (Figure 3B).

    This result highlights another advantage of using metabolically incorporated Photo-Leucine and Photo-Methionine for crosslinking within protein-protein interaction domains: less charge (pI) neutralization of protein side chains and protein polymerization, which could decrease overall protein solubility.


    Conclusion

    Photo-Leucine and Photo-Methionine are unique, protein-specific crosslinking reagents for use in living cells with the ability to capture more diverse protein-protein interactions with better protein recovery after cell lysis than formaldehyde and NHS-ester crosslinkers.

    ---

    Ryan D. Bomgarden, Ph.D., is research scientist at Thermo Fisher Scientific. Web: www.thermo.com/pierce. E-mail: Ryan.Bomgarden@thermofisher.com.



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