Cleavage of peptides and proteins using light-generated radicals from titanium dioxide.
Protein identification and characterization often requires cleavage into distinct fragments. Current methods require proteolytic enzymes or chemical agents and typically a second reagent to discontinue cleavage. We have developed a selective cleavage process for peptides and proteins using light-generated radicals from titanium dioxide. The hydroxyl radicals, produced at the TiO(2) surface using UV light, are present for only hundreds of microseconds and are confined to a defined reagent zone. Peptides and proteins can be moved past the "reagent zone", and cleavage is tunable through residence time, illumination time, and intensity. Using this method, products are observed consistent with cleavage at proline residues. These initial experiments indicate the method is rapid, specific, and reproducible. In certain configurations, cleavage products are produced in less than 10 s. Reproducible product patterns consistent with cleavage of the peptide bond at proline for angiotensin I, Lys-bradykinin, and myoglobin are demonstrated using capillary electrophoresis. Mass characterization of fragments produced in the cleavage of angiotensin I was obtained using liquid chromatography-mass spectrometry. In addition to the evidence supporting cleavage at proline, enkephalin and peptide A-779, two peptides that do not contain proline, showed no evidence of cleavage under the same conditions.
Jones
Barbara J
BJ
Arizona State University, Tempe, Arizona 85257, USA. Barb.jones@nist.gov
Vergne
Matthew J
MJ
Bunk
David M
DM
Locascio
Laurie E
LE
Hayes
Mark A
MA
D019075:Electrophoresis, Capillary; D017665:Hydroxyl Radical / Q000737:chemistry / Q000528:radiation effects; D008027:Light; D010316:Particle Size; D010455:Peptides / Q000737:chemistry / Q000528:radiation effects; D011506:Proteins / Q000737:chemistry / Q000528:radiation effects; D013499:Surface Properties; D013997:Time Factors; D014025:Titanium / Q000737:chemistry* / Q000528:radiation effects
D010455:Peptides; D011506:Proteins; C009495:titanium dioxide; D017665:Hydroxyl Radical; D014025:Titanium