HIGH-RESOLUTION ACCURATE MASS MEASUREMENTS OF BIOMOLECULES USING A NEW ELECTROSPRAY-IONIZATION ION-CYCLOTRON RESONANCE MASS-SPECTROMETER

被引:176
作者
WINGER, BE [1 ]
HOFSTADLER, SA [1 ]
BRUCE, JE [1 ]
UDSETH, HR [1 ]
SMITH, RD [1 ]
机构
[1] PACIFIC NW LAB, DEPT CHEM SCI, CHEM METHODS & SEPARAT GRP,POB 999, MS P8-19, RICHLAND, WA 99352 USA
关键词
D O I
10.1016/1044-0305(93)85018-S
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A novel electrospray ionization/Fourier transform ion cyclotron resonance mass spectrometer based on a 7-T superconducting magnet was developed for high-resolution accurate mass measurements of large biomolecules. Ions formed at atmospheric pressure using electrospray ionization (ESI) were transmitted (through six differential pumping stages) to the trapped ion cell maintained below 10(-9) torr. The increased pumping speed attainable with cryopumping (> 10(5) L/s) allowed brief pressure excursions to above 10(-4) torr, with greatly enhanced trapping efficiencies and subsequent short pumpdown times, facilitating high-resolution mass measurements. A set of electromechanical shutters were also used to minimize the effect of the directed molecular beam produced by the ESI source and were open only during ion injection. Coupled with the use of the pulsed-valve gas inlet, the trapped ion cell was generally filled to the space charge limit within 100 ms. The use of 10-25 ms ion injection times allowed mass spectra to be obtained from 4 fmol of bovine insulin (M(r) 5734) and ubiquitin (M(r) 8565), with resolution sufficient to easily resolve the isotopic envelopes and determine the charge states. The microheterogeneity of the glycoprotein ribonuclease B was examined, giving a measured mass of 14,898.74 Da for the most abundant peak in the isotopic envelope of the normally glycosylated protein (i.e., with five mannose and two N-acetylglucosamine residues (an error of approximately 2 ppm) and an average error of approximately 1 ppm for the higher glycosylated and various H3PO4 adducted forms of the protein. Time-domain signals lasting in excess of 80 s were obtained for smaller proteins, producing, for example, a mass resolution of more than 700,000 for the 4+ charge state (m/z 1434) of insulin.
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收藏
页码:566 / 577
页数:12
相关论文
共 60 条
  • [1] ALEKSANDROV ML, 1984, DOKL AKAD NAUK SSSR, V277, P379
  • [2] BARINAGA CJ, 1989, RAPID COMMUN MASS SP, V3, P160
  • [3] IMPROVED FOURIER-TRANSFORM ION-CYCLOTRON-RESONANCE MASS-SPECTROMETRY OF LARGE BIOMOLECULES
    BEU, SC
    SENKO, MW
    QUINN, JP
    MCLAFFERTY, FW
    [J]. JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 1993, 4 (02) : 190 - 192
  • [4] THE ACCURACY OF HEAVY-ION MASS MEASUREMENTS USING TIME OF FLIGHT-ION CYCLOTRON-RESONANCE IN A PENNING TRAP
    BOLLEN, G
    MOORE, RB
    SAVARD, G
    STOLZENBERG, H
    [J]. JOURNAL OF APPLIED PHYSICS, 1990, 68 (09) : 4355 - 4374
  • [5] AN ION-STORAGE TIME-OF-FLIGHT MASS-SPECTROMETER FOR ANALYSIS OF ELECTROSPRAY IONS
    BOYLE, JG
    WHITEHOUSE, CM
    FENN, JB
    [J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 1991, 5 (09) : 400 - 405
  • [6] TIME-OF-FLIGHT MASS-SPECTROMETRY WITH AN ELECTROSPRAY ION-BEAM
    BOYLE, JG
    WHITEHOUSE, CM
    [J]. ANALYTICAL CHEMISTRY, 1992, 64 (18) : 2084 - 2089
  • [7] PROBING PROTEIN CONFORMATION BY A COMBINATION OF ELECTROSPRAY MASS-SPECTROMETRY AND MOLECULAR MODELING
    BROWN, C
    CAMILLERI, P
    HASKINS, NJ
    SAUNDERS, M
    [J]. JOURNAL OF THE CHEMICAL SOCIETY-CHEMICAL COMMUNICATIONS, 1992, (10) : 761 - 764
  • [8] ON PRODUCTION OF POLARIZED ELECTRON BEAMS BY SPIN EXCHANGE COLLISIONS
    BYRNE, J
    FARAGO, PS
    [J]. PROCEEDINGS OF THE PHYSICAL SOCIETY OF LONDON, 1965, 86 (552P): : 801 - &
  • [9] CARAVATTI P, 1990, Patent No. 4924089
  • [10] CHAIT BT, 1991, 39TH P C MASS SPECTR, P447