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Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of the recombinant human macrophage colony stimulating factor β and derivatives

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Journal of the American Society for Mass Spectrometry

Abstract

The potential of electrospray ionization (ESI) Fourier transform ion cyclotron mass spectrometry (FTICR-MS) to assist in the structural characterization of monomeric and dimeric derivatives of the macrophage colony stimulating factor β (rhM-CSFβ) was assessed. Mass spectrometric analysis of the 49 kDa protein required the use of sustained off-resonance irradiation (SORI) in-trap cleanup to reduce adduction. High resolution mass spectra were acquired for a fully reduced and a fully S-cyanylated monomeric derivative (∼25 kDa). Mass accuracy for monomeric derivatives was better than 5 ppm, after applying a new calibration method (i.e., DeCAL) which eliminates space charge effects upon high accuracy mass measurements. This high mass accuracy allowed the direct determination of the exact number of incorporated cyanyl groups. Collisionally induced dissociation using SORI yielded b- and y-fragment ions within the N- and C-terminal regions for the monomeric derivatives, but obtaining information on other regions required proteolytic digestion, or potentially the use of alternative dissociation methods.

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Authors and Affiliations

  1. Department of Chemistry, Oregon State University, Corvallis, Oregon, USA

    Claudia S. Maier, Xuguang Yan & Max L. Deinzer

  2. Department of Biochemistry & Biophysics, Oregon State University, Corvallis, Oregon, USA

    Mark E. Harder & Michael I. Schimerlik

  3. Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington, USA

    Ljiljana Paša-Tolić & Richard D. Smith

Authors
  1. Claudia S. Maier
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  2. Xuguang Yan
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  3. Mark E. Harder
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  4. Michael I. Schimerlik
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  5. Max L. Deinzer
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  6. Ljiljana Paša-Tolić
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  7. Richard D. Smith
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Correspondence to Max L. Deinzer.

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Maier, C.S., Yan, X., Harder, M.E. et al. Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of the recombinant human macrophage colony stimulating factor β and derivatives. J Am Soc Mass Spectrom 11, 237–243 (2000). https://doi.org/10.1016/S1044-0305(99)00139-7

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  • DOI: https://doi.org/10.1016/S1044-0305(99)00139-7

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