title: Nanoliter chemistry combined with mass spectrometry for peptide mapping of proteins from single mammalian cell lysates.
abstract: A nanoliter-chemistry station combined with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was developed to characterize proteins at the attomole level. Chemical reactions including protein digestion were carried out in nanoliter or subnanoliter volumes, followed by microspot sample deposition of the digest to a MALDI-TOF mass spectrometer. Accurate mass determination of the peptides from the enzyme digest, in conjunction with protein database searching, allowed the identification of the proteins in the protein database. This method is particularly useful for handling small-volume samples such as in single-cell analysis. The high sensitivity and specificity of this method were demonstrated by peptide mapping and identifying hemoglobin variants of sickle cell disease from a single red blood cell. The approach of combining nanoliter chemistry with highly sensitive mass spectrometric analysis should find general use in characterizing proteins from biological systems where only a limited amount of material is available for interrogation.
authors.affiliation: Department of Chemistry, University of Alberta, Edmonton, Canada.
authors.forename: R M
authors.initials: RM
authors.lastname: Whittal
authors.forename: B O
authors.initials: BO
authors.lastname: Keller
authors.forename: L
authors.initials: L
authors.lastname: Li
mesh_terms: D000818:Animals; D008322:Mammals; D010449:Peptide Mapping / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D011506:Proteins