Cleavage of peptides and proteins using light-generated radicals from titanium dioxide

Barbara J. Jones, Matthew J. Vergne, David M. Bunk, Laurie E. Locascio, Mark Hayes

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

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 TiO2 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, Lysbradykinin, 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.

Original languageEnglish (US)
Pages (from-to)1327-1332
Number of pages6
JournalAnalytical Chemistry
Volume79
Issue number4
DOIs
StatePublished - Feb 15 2007

Fingerprint

Proline
Angiotensin I
Peptides
Proteins
Capillary electrophoresis
Myoglobin
Enkephalins
Liquid chromatography
Ultraviolet radiation
Hydroxyl Radical
Mass spectrometry
Peptide Hydrolases
Lighting
titanium dioxide
Experiments

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Cleavage of peptides and proteins using light-generated radicals from titanium dioxide. / Jones, Barbara J.; Vergne, Matthew J.; Bunk, David M.; Locascio, Laurie E.; Hayes, Mark.

In: Analytical Chemistry, Vol. 79, No. 4, 15.02.2007, p. 1327-1332.

Research output: Contribution to journalArticle

Jones, Barbara J. ; Vergne, Matthew J. ; Bunk, David M. ; Locascio, Laurie E. ; Hayes, Mark. / Cleavage of peptides and proteins using light-generated radicals from titanium dioxide. In: Analytical Chemistry. 2007 ; Vol. 79, No. 4. pp. 1327-1332.
@article{f1754594fe4548ceb7eaf51a1d0a9228,
title = "Cleavage of peptides and proteins using light-generated radicals from titanium dioxide",
abstract = "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 TiO2 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, Lysbradykinin, 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.",
author = "Jones, {Barbara J.} and Vergne, {Matthew J.} and Bunk, {David M.} and Locascio, {Laurie E.} and Mark Hayes",
year = "2007",
month = "2",
day = "15",
doi = "10.1021/ac0613737",
language = "English (US)",
volume = "79",
pages = "1327--1332",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "4",

}

TY - JOUR

T1 - Cleavage of peptides and proteins using light-generated radicals from titanium dioxide

AU - Jones, Barbara J.

AU - Vergne, Matthew J.

AU - Bunk, David M.

AU - Locascio, Laurie E.

AU - Hayes, Mark

PY - 2007/2/15

Y1 - 2007/2/15

N2 - 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 TiO2 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, Lysbradykinin, 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.

AB - 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 TiO2 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, Lysbradykinin, 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.

UR - http://www.scopus.com/inward/record.url?scp=33847228064&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33847228064&partnerID=8YFLogxK

U2 - 10.1021/ac0613737

DO - 10.1021/ac0613737

M3 - Article

VL - 79

SP - 1327

EP - 1332

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 4

ER -