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.
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.