Divalent Cation-Mediated Interaction Between Cerebroside Sulfate and Cerebrosides: An Investigation of the Effect of Structural Variations of Lipids by.

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Divalent Cation-Mediated Interaction Between Cerebroside Sulfate and Cerebrosides: An Investigation of the Effect of Structural Variations of Lipids by Electrospray Ionization Mass Spectrometry Kalavelil M. Koshy, Jianyao Wang, Joan M. Boggs Biophysical Journal Volume 77, Issue 1, Pages 306-318 (July 1999) DOI: 10.1016/S0006-3495(99)76891-8 Copyright © 1999 The Biophysical Society Terms and Conditions

Figure 1 ESI mass spectrum of a mixture of HFA-CBS, HFA-GalCer, and calcium chloride in methanol solution in the molar ratio 1:1:10 at a declustering potential of 120V, showing the various ions normally observable under ESI conditions. The compositions of the ions are as follows: 1. [HFA-GalCer.Ca2+]2+ (392.2), 2. [HFA-GalCer.H+]+ (745), 3. [2HFA-GalCer.Ca2+]2+ (764), 4. [HFA-GalCer.Na+]+ (767), 5. [HFA-CBS.Ca2+-H]+ (862.8), 6. [3HFA-GalCer.Ca2+]2+ (1136), 7. [2HFA-CBS.HFA-GalCer.2Ca2+-2H]2+ (1234.8), and 8. [HFA-CBS.HFA-GalCer.Ca2+-H]+ (1606.4). The numbers in parentheses are the observed m/z values. Biophysical Journal 1999 77, 306-318DOI: (10.1016/S0006-3495(99)76891-8) Copyright © 1999 The Biophysical Society Terms and Conditions

Figure 2 ESI mass spectra of a mixture of NFA-CBS, NFA-GalCer-d35, NFA-GlcCer, and calcium chloride in the ratio 2:1:1:10 at declustering potentials of 100V (A), 150V (B), and 180V (C). The peaks of interest are the following: 1. A set of three peaks corresponding to the compositions [2NFA-GlcCer.Ca2+]2+ (747.6), [NFA-GlcCer.NFA-GalCer-d35.Ca2+]2+ (765), and [2NFA-GalCer-d35.Ca2+]2+ (783) (peaks at 751 and 786 are the sodium adducts of NFA-GlcCer and NFA-GalCer-d35, respectively); 2. [NFA-CBS.Ca2+-H]+ (847); 3. a set of three peaks corresponding to the compositions [3NFA-GlcCer.Ca2+]2+ (1111.4), [2NFA-GlcCer.NFA-GalCer-d35.Ca2+]2+ (1129.4), and [NFA-GlcCer.2NFA-GalCer-d35.Ca2+]2+ (1146.4); 4. a set of two peaks corresponding to the compositions [2NFA-CBS.GlcCer.2Ca2+-2H]2+ (1211.0) and [2NFA-CBS.GalCer-d35.2Ca2+-2H]2+ (1229.0); 5. a set of two peaks corresponding to the compositions [NFA-CBS.NFA-GlcCer.Ca2+-H]+ (1574.8) and [NFA-CBS.NFA-GalCer-d35.Ca2+-H]+ (1609.4). Biophysical Journal 1999 77, 306-318DOI: (10.1016/S0006-3495(99)76891-8) Copyright © 1999 The Biophysical Society Terms and Conditions

Figure 3 The variation of the relative intensity of the peak due to the heterodimeric ion [HFA-CBS.GCer.Zn2+-H]+ with declustering potential. □, NFA-GalCer; ♢, HFA-GalCer; ○, NFA-GlcCer; △, HFA-GlcCer. Biophysical Journal 1999 77, 306-318DOI: (10.1016/S0006-3495(99)76891-8) Copyright © 1999 The Biophysical Society Terms and Conditions

Figure 4 ESI mass spectrum of a mixture of HFA-CBS, HFA-GalCer, and zinc chloride in methanol in the molar ratio 1:1:10 at a declustering potential of 180V. The peaks indicated are: 1. [HFA-GalCer.Zn2+-H-H2O]+ (788.2), 2. [HFA-GalCer.Zn2+-H]+ (806.8), 3. [HFA-CBS.Zn2+-H]+ (886.8), and 4. [HFA-CBS.HFA-GalCer.Zn2+-H]+ (1632.4). See footnote to Table 4. The numbers in parentheses are the m/z values. Biophysical Journal 1999 77, 306-318DOI: (10.1016/S0006-3495(99)76891-8) Copyright © 1999 The Biophysical Society Terms and Conditions

Figure 5 ESI mass spectrum of a mixture of galactose 3-sulfate, galactose, and calcium chloride in the molar ratio of 1:1:10, in acetonitrile/water mixture (50:50) at a declustering potential of 100V. Peaks indicated are 1. [Gal.Ca2+-H]+ (218.8), 2. [Gal.Ca2+.Cl−]+ (254.8), 3. [GalS.Ca2+-H]+ (298.8), 4. [2Gal.Ca2+-H]+ (398.8), 5. [2Gal.Ca2+.Cl−]+ (434.8), and 6. [GalS.Gal.Ca2+-H]+ (479.0). The numbers in parentheses are the m/z values. Biophysical Journal 1999 77, 306-318DOI: (10.1016/S0006-3495(99)76891-8) Copyright © 1999 The Biophysical Society Terms and Conditions

Figure 6 ESI mass spectrum of a mixture of acetylated lysosulfatide, β-methylgalactoside, and calcium chloride in the mole ratio 1:10:10 in acetonitrile/water mixture (50:50) at a declustering potential of 60V, showing the [AcLysCBS.Ca2+-H]+ ion, peak 1 (622) and [AcLysCBS.MeGal.Ca2+-H]+, peak 2 (816.4). The origin of peak 3 (904) is obscure. Although this peak appeared in the spectrum of both β-methyl galactoside and β-methyl glucoside individually, in the competition experiment using β-methyl galactoside-d3 and β-methyl glucoside, this peak was not observed. CID experiments showed that [AcLysCBS.Ca2+-H]+ is a component of this ion. Biophysical Journal 1999 77, 306-318DOI: (10.1016/S0006-3495(99)76891-8) Copyright © 1999 The Biophysical Society Terms and Conditions