Fishing for Membrane Proteins Using the Synthesized CB[7] & AFc Binding Pair

Mentors: Thomas Lau, Jennifer Beaudoin, Eamon Comer

Plasma membrane proteins play crucial roles in countless biological processes. To study them, an efficient technique for extracting, purifying, and identifying membrane proteins is necessary. Membrane proteins are difficult to extract, however, because of their hydrophobicity and microheterogeneity. Soun investigated if a new binding pair – the molecule CB[7] and a Ferrocene (Fc) derivative compound – might be used for a membrane protein extraction technique, to obtain high yields and levels of purity.

To test out this new method, Soun synthesized the CB[7] molecule and loaded it onto beads. He also synthesized the Ferrocene derivative Afc, and bound Afc to all of the membrane proteins in a human breast cancer cell line. He then lysed the breast cancer cells, and used the CB[7]-loaded beads to capture all of the membrane proteins in these cells. Proteins were eluted and digested with trypsin, and the resulting peptides were identified by LC-MS. The mass spec data will be used to determine how the new capturing method compares to the standard methods, such as those using biotin and streptavidin.



Soun, a senior at Revere High School, tested a new technique for extracting membrane proteins from cells, with the goal of increasing yield and purity, compared to existing methods.