Abstract
Ultra-high-sensitivity, biopolymer sequencing is a goal in many fields of molecular biology, and collisionally activated decomposition electrospray mass spectrometry (CAD ES MS/MS) using a triple quadrupole mass spectrometer has become a method of choice for work in the high- to mid-femtomole range. However, when the detection of ions becomes statistical, as it may in that range, the mass assignment of fragment ions is inaccurate and either sequencing becomes impossible or ambiguities result due, for example, to the closeness in amino acid residue masses (I/L, N or K/Q, E). Some ambiguities may be resolved by synthesizing possible sequences, but this is unsatisfactory. In considering the limitations of triple quadrupole MS/MS with respect to scanning ion detection, resolution, transmission, and mass accuracy, we reasoned that a novel geometry quadrupole orthogonal acceleration time-of-flight (Q-TOF) instrument would have special merit for ultra-high-sensitivity MS/MS sequencing, and suggested its construction for this purpose some three years ago. A prototype Q-TOF has now been built by Micromass [Morris et al. (1996), Rapid Commun. Mass Spectrom. 10, 889–896], and in the first research on the instrument, including MHC antigen and filarial nematode glycoprotein studies, we demonstrate low-femtomole- and attomole-range sequencing with mass accuracy of better than 0.1 Da throughout the daughter-ion spectrum, thus removing sequencing ambiguities in some of the most challenging work demanding the highest sensitivity.
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REFERENCES
Batley, K. E., and Morris, H. R. (1977a). Biochem. Biophys. Res. Commun. 75, 1010–1014.
Batley, K., and Morris, H. R. (1977b). Biochem. Soc. Trans. 5, 1097–1100.
Beloff-Chain, A., Morton J., Dunmore S., Taylor G. W., and Morris H. R. (1983). Nature 301, 255–258.
Brain S. D., Williams, T. J., Tippins J. R., Morris H. R., and MacIntyre I. (1984). Nature 313, 54–56.
Bridgen J., and Morris, H. R. (1974). Eur. J. Biochem. 44, 333–334.
Chan, A. L., Morris, H. R., Panico, M., Etienne, A., Rogers, M. E., Gaffney P., Creighton-Kempsford, L., and Dell, A. (1991). Glycobiology, 1, 173–186.
Dell, A., Morris, H. R., Easton, R. L., Panico, M., Patankar, M., Oehninger, S., Koistinen, R., Koistinen, H., Seppala, M., and Clark, G. F. (1995). J. Biol. Chem., 270, 24116–24126.
Erdjument, D. A., Lane, D. A., Panico, M., Di Marzo, V., and Morris, H. R. (1988). J. Biol. Chem. 263, 5589–5593.
Geddes, A. J., Graham, G. N., Morris, H. R., Lucas, F., Barber, M., and Wolstenholme, W. A. (1969). Biochem. J. 114, 695–702.
Hughes, J., Smith, T. W., Kosterlitz, H. W., Fothergill, L., Morgan, B. A., and Morris, H. R. (1975). Nature 258, 577–579.
Hunt, D. F., Michel, H., Dickinson, T. A., Shabanowitz, J., et al. (1992). Science 256, 1817–1820.
Khoo, K.-H., Sarda, S., Xa, X., Caulfield, J. P., McNeil, M. R., Homans, S. W., Morris, H. R., and Dell, A. (1995). J. Biol. Chem., 270, 17114–17123.
Magnusson, S., Sottrup-Jensen, L., Petersen, T. E., Morris, H. R., and Dell, A. (1974). FEBS Lett. 44, 189–193.
Morris, H. R. (1972). FEBS Lett. 22, 257–260.
Morris, H. R., and Dell, A. (1975). Biochem. J. 149, 754–755.
Morris, H. R., and Williams, D. H. (1972). J. Chem. Soc. Chem. Commun. 1972, 114–116.
Morris, H. R, Geddes, A. J., and Graham, G. N. (1969). Biochem. J. 111, 38.
Morris, H. R., Williams, D. H., and Ambler, R. P. (1971). Biochem. J. 125, 189–201.
Morris H. R., Batley, K. E., Harding, N. G. L., Bjur, R. A., Dann, J. G., and King, R. W. (1974a). Biochem. J. 137, 409–411.
Morris, H. R., Williams, D. H., Midwinter, G. C., and Hartley, B. S. (1974b). Biochem. J. 141, 701–713.
Morris, H. R., Dell, A., Petersen, T. E., Sottrup-Jensen, L., and Magnusson, S. (1976). Biochem. J. 153, 663–679.
Morris, H. R., Thompson, M. R., Osuga, D. T., Ahmed, A. I., Chan, S. M., Vandenheede, J., and Feeney R. (1978). J. Biol. Chem., 253, 5155–5161.
Morris, H. R., Taylor, G. W., Piper, P. J., Tippins, J. R., and Smahoun, M. N. (1980). Nature, 285, 104–106.
Morris, H. R., Panico, M., Karplus, A., Lloyd, P. E., and Riniker, B. (1982). Nature 300, 643–645.
Morris, H. R., Panico, M., Etienne, A. T., MacIntyre, I., Tippins, J. R., and Girgis, S. I. (1984). Nature 308, 746–748.
Morris, H. R., McDowell, R., Dell, A., and Panico, M. (1991). In Proceedings of the 39th American Society for Mass Spectrometry Conference on MS and Allied Topics, Nashville, pp. 1673–1674.
Morris, H. R., Dell, A., Panico, M., and McDowell, R., (1995). In Proceedings of the 43rd American Society for Mass Spectrometry Conference on MS and Allied Topics, Atlanta, p. 360.
Morris, H. R., Morris, H. R., Paxton, T., Dell, A., Langhorne, J., Berg, M., Bordoli, R. S., Hoyes, J., and Bateman R. H. (1996a). Rapid Commun. Mass Spectrom. 10, 889–896
Morris, H. R., Dell, A., Easton, R. L., Panico, M., Koistinen, H., Koistinen, R., Oehninger, S., Patankar, M. S., Seppala, M., and Clark, G. F. (1996b). J. Biol. Chem. 271, 32159–32167.
Reason, A. J., Treisman, R. H., Marais, R., Haltiwanger, R. S., Hart, G. W., Morris, H. R., Panico, M., and Dell, A. (1992). J. Biol. Chem. 267, 16911–16921.
Robb, R. J., Kutney, R. M., Panico, M., Morris, H. R., De Grado, W. F., and Chowdry, V. (1983). Biochem. Biophys. Res. Commun. 116, 1049–1055.
Robb, R. J., Kutney, R. M., Panico, M., Morris, H. R., De Grado, W. F., and Chowdry V. (1984). Proc. Natl. Acad. Sci. USA 81, 6486–6490.
Sasaki, H., Bothner, B., Dell, A., and Fukuda, M. (1987). J. Biol. Chem. 262, 12059–12076.
Shaw, W. V., Packman, L. C., Burghleigh, B. D., Dell, A., Morris, H. R., and Hartley, B. S. (1979). Nature 282, 870–872.
Spooncer, E., Fukuda, M., Klock, J. C., Oates, J. E., and Dell, A. (1984). J. Biol. Chem., 259, 4792–4801.
Stone, J. V., Mordue, W., Batley, K. E., and Morris, H. R. (1976). Nature 263, 207–211.
Thogersen, H. C., Petersen, T. E., Sottrup-Jensen, L., Magnusson, S., and Morris, H. R. (1978). Biochem. J. 175, 613–627.
Thomas, D. W., Das, B. C., Gero, S. D., and Lederer, E. (1968). Biochem. Biophys. Res. Commun 32, 199–202.
Wang, W., Meadows, L. R., den Haan, J. M. M. et al. (1995). Science 269, 1588–1590.
Williamson, R. A., Marston, F. A. O., Angal, S., Koklitis, P., Panico, M., Morris, H. R., Carne, A. F., Smith, B. J., Harris, T. J. R., and Freedman, R. B. (1990). Biochem J. 268, 267–274.
Wilm, M., Schevchenko, A., Houthaeve, T., Breit, S., et al. (1996) Nature 379, 466–469.
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Morris, H.R., Paxton, T., Panico, M. et al. A Novel Geometry Mass Spectrometer, the Q-TOF, for Low-Femtomole/Attomole-Range Biopolymer Sequencing. J Protein Chem 16, 469–479 (1997). https://doi.org/10.1023/A:1026309410737
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DOI: https://doi.org/10.1023/A:1026309410737