High-Mass-Measurement Accuracy and 100 Sequence Coverage of Enzymatically Digested Bovine Serum Albumin from an ESI-FTICR Mass Spectrum
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- 作者：James E. Bruce ; Gordon A. Anderson ; Jenny Wen ; Richard Harkewicz ; and Richard D. Smith
- 刊名：Analytical Chemistry
- 出版年：1999
- 出版时间：July 15, 1999
- 年：1999
- 卷：71
- 期：14
- 页码：2595 - 2599
- 全文大小：49K
- 年卷期：v.71,no.14(July 15, 1999)
- ISSN：1520-6882
文摘
The application of Fourier transform ion cyclotron resonance (FTICR) mass spectrometry to the analysis ofpolypeptide mixtures resulting from proteolytic digestionis described. A new 11.5-T FTICR mass spectrometer hasbeen applied for the analysis of tryptic digestion mixturesof the protein bovine serum albumin (BSA). The improvedcyclotron frequency stability and reduced frequency shiftsobserved over a wide range of trapped ion population sizesprovide the ability to signal average spectra withoutdegrading mass measurement accuracy, requiring internal calibration or advanced data processing schemes tocompensate for variations in ion cyclotron signals broughtabout by different population sizes. A total of 100 spectrawere signal-averaged leading to the observation of a totalof 123 isotope distributions with a signal-to-noise ratiogreater than 3:1. From those distributions, 86 can beascribed to tryptic fragments of BSA on the basis of massmeasurement errors of 10 ppm or less. Of these, 71 werewithin 2 ppm error limits corresponding to completeamino acid sequence coverage and an average error of0.77 ppm. These results indicate that high-accuracymeasurements are feasible for a large number of speciesdetected simultaneously without the necessity for internalcalibration and indicate the potential of such measurements, when combined with chromatographic separations, for facilitating more rapid identification of largenumbers of proteins.