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Blood, Vol. 113, Issue 11, 2578-2586, March 12, 2009
Haptoglobin preserves the CD163 hemoglobin scavenger pathway by shielding hemoglobin from peroxidative modification
Blood Buehler et al.
113: 2578
Supplemental materials for: Buehler et al
Files in this Data Supplement:
- Document 1. Supplemental materials, methods, and results (PDF, 76.8 KB)
- Table S1. Monisotopic tryptic peptide mass changes in hemoglobin after oxidation (PDF, 63.2 KB)
- Table S2. Haptoglobin binding region distances from protected amino acids (PDF, 63.2 KB)
- Figure S1.Collision-induced dissociation spectra of Hb peptides (βTrp15 and βMet55) (JPG, 118 KB)
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(A) oxidized HbM2H2 ion at m/z468.7597 corresponding to the tryptic peptide at β9–17 containing oxidized Trp-15 found in H2O2 treated samples. (B) Oxidized Hb-Hp -M2H2 ion at m/z474.7599 corresponding to the tryptic peptide at β9–17 containing intact Trp-15 found in untreated samples. (C) M2H2 ion at m/z1037.97 for Met-55 sulfoxide corresponding to the tryptic peptide at β41–59 containing Met-55 both treated and untreated samples. The resulting N-terminal generated b ions and C-terminal generated y ions are labeled accordingly in the spectra.
- Figure S2. Collision-induced dissociation spectra of Hb peptides (βCys93 and βCys112) (JPG, 136 KB)
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(A) M2H2 ion at m/z735.33 corresponding to the tryptic peptide at β93–105 containing oxidized Cys-93 found in H2O2 treated samples. (B) MH1 ion at m/z2586.33 corresponding to the of the tryptic peptide at β93–114 containing intact Cys-93 found in untreated samples. (C) M3H3 ion at m/z589.99 corresponding to the of the tryptic peptide at β105–120 containing oxidized Cys-112 found in H2O2 treated samples. (D) M3H3 ion at m/z593.00 corresponding to the tryptic peptide at β105–120 containing oxidized Cys-112 found in untreated samples. The resulting N-terminal generated b ions and C-terminal generated y ions are labeled in the spectra.
- Figure S3. Collision-induced dissociation spectra of histidine containing peptides (JPG, 93.3 KB)
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(A) M3H3+ ion at m/z598.7 corresponding to the of the tryptic peptide at 105-120, corresponding to mass of 1792.076, containing oxidized His 117 found in H2O2 treated samples. (B) M3H3+ ion at m/z494.6 corresponding to the of the tryptic peptide ( at 133-146, corresponding to mass of 1480.8, containing oxidized His-15 and Tyr-14 found in H2O2 treated samples. The resulting N-terminal generated b ions and C-terminal generated y ions are labeled in the spectra.
- Figure S4. Proximity of irreversibly oxidizable Hb β chain amino acids to Hp binding regions in ribbon representation of the Hb α-β dimmer (JPG, 62.7 KB)
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(A) Blue - represents the Hb dimer β globin chains, Red – represents the Hb dimer α globin chains, green ball and stick represent heme moieties. Within the β chains (blue), two amino acid regions indicated in yellow (β 11-25 and β 134-146) 27, 31 represent defined Hp binding regions. Within the α chain (red), one amino acid region indicated in yellow (α 121-135) 29 represents a defined region for Hp binding. Distance measurements were made using PDB Protein Workshop 1.50. Calculations were made from each irreversibly oxidizable amino acid represented as space filling atoms (B) βCYS93, (C) βCYS112 and (D) βTRP15 to the C-terminal, N- terminal and midpoint of literature defined Hp binding regions within the Hb β and α chains. Distances of all oxidizable amino acids to the defined regions of Hp binding are included in the supplemental material.
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