Photo-Induced Cross-Linking of Unmodified Proteins (PICUP) is a protein cross-linking method by visible light irradiation of a photocatalyst in the presence of an electron acceptor and the protein of interest. Irradiation results in a highly reactive protein radical that forms a covalent bond between the amino acid side chains of the proteins to be linked. Cross-linking methods developed prior to PICUP, including the use of physical, oxidative, and chemical cross-linkers, often require more time and result in protein byproducts. In addition, the cross-linked protein yield is very low due to the multifunctionality of the cross-linking reagents.
The process was invented (US6613582B1) in 1999 to utilize protein cross-linking techniques to analyze the interactions between polypeptides as well as structural differences proteins undergo in a catalytic pathway. The techniques in the 20th century were not sufficient to be applied to cross-link fast and transient changes of these proteins in high yield. PICUP allowed for rapid (<1 second) and high production of covalently-linked proteins in close proximity with each other.