Capture of phosphopeptides using alpha-zirconium phosphate nanoplatelets.
Journal Article
Alpha-zirconium phosphate nanoplatelets (alpha-ZrPN) were studied as a binding agent for phosphopeptides. Nanoplatelets of alpha-zirconium phosphate were incubated overnight with zirconium oxychloride, followed by centrifugation, and washed twice with water followed by an aqueous solution of 80% acetonitrile to form the binding agent. Alpha-ZrPN were able specifically to capture phosphoserine-containing peptides from a tryptic digest of a complex peptide mixture in which its abundance was only 0.05%. Alpha-ZrPN also bound peptides containing phosphothreonine and phosphotyrosine. The limit of detection for phosphopeptides is approximately 2 fmol, based on using matrix-assisted laser desorption/ionization mass spectrometry. Alpha-ZrPN were applied for the analysis of tryptic digests of mouse liver and leukemia cell phosphoproteomes and succeeded in identifying 158 phosphopeptides (209 phosphorylation sites) from 101 phosphoproteins in mouse liver lysate and 78 phosphopeptides (104 phosphorylation sites) from 59 phosphoproteins in leukemia cell extract. For these two tryptic digests, the alpha-ZrPN approach is able to capture more phosphopeptides than that obtained from TiO2 particles or from Fe(3+)-IMAC beads, but each method is able to bind some phosphopeptides that the others do not.
Full Text
Duke Authors
Cited Authors
- Xu, S; Whitin, JC; Yu, TT-S; Zhou, H; Sun, D; Sue, H-J; Zou, H; Cohen, HJ; Zare, RN
Published Date
- July 15, 2008
Published In
Volume / Issue
- 80 / 14
Start / End Page
- 5542 - 5549
PubMed ID
- 18522436
Pubmed Central ID
- 18522436
Electronic International Standard Serial Number (EISSN)
- 1520-6882
Digital Object Identifier (DOI)
- 10.1021/ac800577z
Language
- eng
Conference Location
- United States