In situ enrichment of phosphopeptides on MALDI plates functionalized by reactive landing of zirconium(IV)-n-propoxide ions.
We report substantial in situ enrichment of phosphopeptides in peptide mixtures using zirconium oxide coated plates for detection by MALDI-TOF mass spectrometry. The novel feature of this approach rests on the specific preparation of zirconium oxide coatings using reactive landing on stainless steel support of gas-phase positive ions produced by electrospray of zirconium(IV)-n-propoxide solutions in 1-propanol. Reactive landing was found to produce durable functionalized surfaces for selective phosphopeptide capture and desorption-ionization by MALDI. Enrichment factors on the order of 20-90 were achieved for several monophosphorylated peptides relative to abundant nonphosphorylated peptides in tryptic digests. We demonstrate the ability of the zirconium oxide functionalized MALDI surfaces to facilitate detection of enriched phosphopeptides in mid-femtomole amounts of alpha-casein digests per MALDI spot.
Blacken
Grady R
GR
Departments of Chemistry, University of Washington, Box 351700, Seattle, Washington 98195-1700, USA.
Volný
Michael
M
Vaisar
Tomás
T
Sadílek
Martin
M
Turecek
Frantisek
F
D000433:1-Propanol / Q000737:chemistry; D000595:Amino Acid Sequence; D002364:Caseins / Q000737:chemistry; D007477:Ions / Q000737:chemistry; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000032:analysis*; D012996:Solutions / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013193:Stainless Steel / Q000737:chemistry*; D013499:Surface Properties; D014357:Trypsin / Q000737:chemistry; D015040:Zirconium / Q000737:chemistry*
D002364:Caseins; D007477:Ions; D010748:Phosphopeptides; D012996:Solutions; D013193:Stainless Steel; D000433:1-Propanol; D015040:Zirconium; D014357:Trypsin; C028541:zirconium oxide