Dynamics and reactivity of HbXL99 alpha. A cross-linked hemoglobin derivative

• Published in: The Journal of biological chemistry 1990-03, Vol.265 (8), p.4449-4454
• Main Authors: Larsen, R W, Chavez, M D, Ondrias, M R, Courtney, S H, Friedman, J M, Lin, M J, Hirsch, R E
• Format: Article
• Language: English
• Subjects: Aspirin - analogs & derivatives; Carbon Monoxide; Cross-Linking Reagents; Fluorescence; Hemoglobin A; Hemoglobins; Hydrogen-Ion Concentration; Photolysis; Raman spectroscopy; Spectrum Analysis, Raman; Tryptophan
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)39585-7

Resonance Raman spectroscopy, transient absorption, and fluroescence techniques have been employed to investigate the structure and dynamics of the alpha-cross-linked hemoglobin derivative, HbXL99 alpha. The resonance Raman spectra of the deoxy form of HbXL99 alpha are identical to those of native NbA (VFe-His approximately 222 cm-1), which exhibit a T-state (low affinity) structure regardless of solvent conditions. The resonance Raman spectra of the transient heme photoproduct resulting from CO photolysis from HbXL99 alpha appear to have structures intermediate between deoxy-T and ligand-bound R structures (VFe-His approximately 222 cm-1). Time-resolved resonance Raman data of HbXL99 alpha-CO show that complete CO recombination occurs after approximately 5 ms, with only a small amount of the CO-bound species reforming within approximately 200 ns (geminate recombination). Transient absorption spectra of HbXL99 alpha-O2 indicate that the extent of sub-nanosecond geminate recombination of O2 is also reduced in the cross-linked derivative relative to native HbA. The decrease in tryptophan fluorescence of HbXL99 alpha upon oxygenation further indicates that tertiary structural changes at the alpha 1-beta 2 interface upon ligation are apparently reduced, but not eliminated in the cross-linked derivative relative to HbA.