Optimizing TiO2-based phosphopeptide enrichment for automated multidimensional liquid chromatography coupled to tandem mass spectrometry.
An automated online multidimensional liquid chromatography system coupled to ESI-based tandem mass spectrometry was used to assess the effectiveness of TiO2 in the enrichment of phosphopeptides from tryptic digests of protein mixtures. By monitoring the enrichment of phosphopeptides, an optimized set of loading, wash, and elution conditions were realized for TiO2. A comparison of TiO2 with other resins used for phosphopeptide enrichment, Fe(III)-IMAC and ZrO2, was also carried out using tryptic digests of both simple and moderately complex protein mixtures; where TiO2 was shown to be superior in performance.
Cantin
Greg T
GT
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, SR11, La Jolla, California 92037, USA.
Shock
Teresa R
TR
Park
Sung Kyu
SK
Madhani
Hiten D
HD
Yates
John R
JR
D000595:Amino Acid Sequence; D002364:Caseins / Q000378:metabolism; D002412:Cations; D002853:Chromatography, Liquid / Q000379:methods*; D007093:Imidazoles / Q000737:chemistry; D007501:Iron / Q000737:chemistry; D008969:Molecular Sequence Data; D009942:Organometallic Compounds / Q000737:chemistry; D010748:Phosphopeptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014025:Titanium / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism; D015040:Zirconium / Q000737:chemistry
D002364:Caseins; D002412:Cations; D007093:Imidazoles; D009942:Organometallic Compounds; D010748:Phosphopeptides; C009495:titanium dioxide; D015040:Zirconium; D014025:Titanium; D007501:Iron; D014357:Trypsin; C005954:imidazoleacetic acid; C028541:zirconium oxide