Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometric analysis of the recombinant human macrophage colony stimulating factor β and derivatives
| Autor: | Ljiljana Paša-Tolić, Richard D. Smith, Claudia S. Maier, Max L. Deinzer, Xuguang Yan, Michael I. Schimerlik, Mark E. Harder |
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| Rok vydání: | 2000 |
| Předmět: |
Protein Folding
Protein mass spectrometry Electrospray ionization Molecular Sequence Data Analytical chemistry Ion cyclotron resonance spectrometry Mass spectrometry Top-down proteomics Mass Spectrometry Fourier transform ion cyclotron resonance Structural Biology Escherichia coli Humans Amino Acid Sequence Chromatography High Pressure Liquid Spectroscopy Chromatography Fourier Analysis Chemistry Granulocyte-Macrophage Colony-Stimulating Factor Cyclotrons Peptide Fragments Recombinant Proteins Mass spectrum Oxidation-Reduction Ion cyclotron resonance |
| Zdroj: | Journal of the American Society for Mass Spectrometry. 11:237-243 |
| ISSN: | 1044-0305 |
| Popis: | 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 beta (rhM-CSF beta) 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 (approximately 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. |
| Databáze: | OpenAIRE |
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