ESI/ion trap/ion mobility/time-of-flight mass spectrometry for rapid and sensitive analysis of biomolecular mixtures

被引:182
作者
Henderson, SC [1 ]
Valentine, SJ [1 ]
Counterman, AE [1 ]
Clemmer, DE [1 ]
机构
[1] Indiana Univ, Dept Chem, Bloomington, IN 47405 USA
关键词
D O I
10.1021/ac9809175
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
An ion trap/ion mobility/time-of-flight mass spectrometry technique is shown to be a rapid and sensitive means of analyzing peptide/protein mixtures. In this approach, an ion trap is used to accumulate ions that have been electrosprayed from a mixture into concentrated packets. The ion packets are injected into a drift tube where components of the mixture are separated based on differences in mobility through a buffer gas. Ions that exit the drift tube are dispersed in a time-of-flight mass spectrometer for mass-to-charge (m/z) determination. The gas-phase separation strategy reduces congestion in the mass spectrum, and experimental mobilities complement m/z measurements in assigning peaks. Examples of the application of the approach to identification of peptides (from tryptic digests) and to separation of charge-state distributions from electrospray of a mixture containing ubiquitin and myoglobin are presented. Most peptides that are observed from tryptic digests of proteins such as cytochrome c and myoglobin can be identified from data that are acquired in under 1 min; studies of mixtures with known compositions indicate that detection limits are similar to 0.5-3 pmol for individual components. Factors that may influence the distributions that are observed, such as storage time in the trap, injection voltages used for the mobility experiment, and variations in ion cross section with charge state, are discussed.
引用
收藏
页码:291 / 301
页数:11
相关论文
共 68 条
  • [1] CHARACTERIZATION BY TANDEM MASS-SPECTROMETRY OF STRUCTURAL MODIFICATIONS IN PROTEINS
    BIEMANN, K
    SCOBLE, HA
    [J]. SCIENCE, 1987, 237 (4818) : 992 - 998
  • [2] BIEMANN K, 1992, ANNU REV BIOCHEM, V61, P997
  • [3] *BIOSYM MSI, 1995, INS 2
  • [4] BURLINGAME AL, 1994, ANAL CHEM, V66, pR634
  • [5] WEIGHING NAKED PROTEINS - PRACTICAL, HIGH-ACCURACY MASS MEASUREMENT OF PEPTIDES AND PROTEINS
    CHAIT, BT
    KENT, SBH
    [J]. SCIENCE, 1992, 257 (5078) : 1885 - 1894
  • [6] PROBING CONFORMATIONAL-CHANGES IN PROTEINS BY MASS-SPECTROMETRY
    CHOWDHURY, SK
    KATTA, V
    CHAIT, BT
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1990, 112 (24) : 9012 - 9013
  • [7] Clemmer DE, 1997, J MASS SPECTROM, V32, P577
  • [8] Space-velocity correlation focusing
    Colby, SM
    Reilly, JP
    [J]. ANALYTICAL CHEMISTRY, 1996, 68 (08) : 1419 - 1428
  • [9] QUADRUPOLE ION TRAP MASS-SPECTROMETRY
    COOKS, RG
    KAISER, RE
    [J]. ACCOUNTS OF CHEMICAL RESEARCH, 1990, 23 (07) : 213 - 219
  • [10] A 2ND GENERATION FORCE-FIELD FOR THE SIMULATION OF PROTEINS, NUCLEIC-ACIDS, AND ORGANIC-MOLECULES
    CORNELL, WD
    CIEPLAK, P
    BAYLY, CI
    GOULD, IR
    MERZ, KM
    FERGUSON, DM
    SPELLMEYER, DC
    FOX, T
    CALDWELL, JW
    KOLLMAN, PA
    [J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1995, 117 (19) : 5179 - 5197