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Evaluating the capacity to generate and preserve nitric oxide bioactivity in highly purified earthworm erythrocruorin: a giant polymeric hemoglobin with potential blood substitute properties.

Roche CJ, Talwar A, Palmer AF, Cabrales P, Gerfen G, Friedman JM - J. Biol. Chem. (2014)

Bottom Line: A potentially important additional property is the capacity of the HBOC either to generate nitric oxide (NO) or to preserve NO bioactivity to compensate for decreased levels of NO in the circulation.The results show that LtHb undergoes the same reactions as HbA and that the reduced efficacy for these reactions for LtHb relative to HbA is consistent with the much higher redox potential of LtHb.Evidence of functional heterogeneity in LtHb is explained in terms of the large difference in the redox potential of the isolated subunits.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, New York 10461.

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Related in: MedlinePlus

Changes in the DAF-FM fluorescence emission for LtHb solutions. The increase and the shift in the emission spectrum of DAF-FM with time when the fluorophore (5 μm final concentration) was added to a solution of LtHb (0.3 mm) are shown. A, metLtHb to which was added nitrite and NO. Black line, initial emission spectrum upon addition of 5 μm DAF to the LtHb solution; red line, change in the emission after 20 min; blue line, change in emission after 40 min. B, a control ferrous NO sample in the absence of nitrite as described under “Experimental Procedures.” Black line, initial emission spectrum upon addition of 5 μm DAF; red line, change in the emission after 20 min; blue line, change in emission after 40 min. C, the evolution of the population distribution profile for the sample prepared to promote the formation of the intermediate used with DAF-FM as described under “Experimental Procedures.” D, the comparable evolution of the population distribution profile for the control sample to which DAF-FM was added. inter, intermediate.
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Figure 12: Changes in the DAF-FM fluorescence emission for LtHb solutions. The increase and the shift in the emission spectrum of DAF-FM with time when the fluorophore (5 μm final concentration) was added to a solution of LtHb (0.3 mm) are shown. A, metLtHb to which was added nitrite and NO. Black line, initial emission spectrum upon addition of 5 μm DAF to the LtHb solution; red line, change in the emission after 20 min; blue line, change in emission after 40 min. B, a control ferrous NO sample in the absence of nitrite as described under “Experimental Procedures.” Black line, initial emission spectrum upon addition of 5 μm DAF; red line, change in the emission after 20 min; blue line, change in emission after 40 min. C, the evolution of the population distribution profile for the sample prepared to promote the formation of the intermediate used with DAF-FM as described under “Experimental Procedures.” D, the comparable evolution of the population distribution profile for the control sample to which DAF-FM was added. inter, intermediate.

Mentions: Figs. 11 and 12 show the evolution of the DAF-FM fluorescence spectra under anaerobic conditions when DAF-FM was added to samples prepared as either the ferrous NO derivative or the NA intermediate of HbA and LtHb. The absorption spectrum was used to establish that in each case the populations were overwhelmingly either ferrous NOHb or the intermediate. For both HbA and LtHb, the increase in fluorescence intensity and the red shift of the fluorescence peak were greater and more rapid for the sample that manifests the absorption spectrum attributable to the NA intermediate (nitrite and NO both added). A similar shift pattern was seen in the absorption maximum of DAF-FM that is also indicative of triazole formation for the samples manifesting the NA intermediate spectrum (not shown).


Evaluating the capacity to generate and preserve nitric oxide bioactivity in highly purified earthworm erythrocruorin: a giant polymeric hemoglobin with potential blood substitute properties.

Roche CJ, Talwar A, Palmer AF, Cabrales P, Gerfen G, Friedman JM - J. Biol. Chem. (2014)

Changes in the DAF-FM fluorescence emission for LtHb solutions. The increase and the shift in the emission spectrum of DAF-FM with time when the fluorophore (5 μm final concentration) was added to a solution of LtHb (0.3 mm) are shown. A, metLtHb to which was added nitrite and NO. Black line, initial emission spectrum upon addition of 5 μm DAF to the LtHb solution; red line, change in the emission after 20 min; blue line, change in emission after 40 min. B, a control ferrous NO sample in the absence of nitrite as described under “Experimental Procedures.” Black line, initial emission spectrum upon addition of 5 μm DAF; red line, change in the emission after 20 min; blue line, change in emission after 40 min. C, the evolution of the population distribution profile for the sample prepared to promote the formation of the intermediate used with DAF-FM as described under “Experimental Procedures.” D, the comparable evolution of the population distribution profile for the control sample to which DAF-FM was added. inter, intermediate.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4281771&req=5

Figure 12: Changes in the DAF-FM fluorescence emission for LtHb solutions. The increase and the shift in the emission spectrum of DAF-FM with time when the fluorophore (5 μm final concentration) was added to a solution of LtHb (0.3 mm) are shown. A, metLtHb to which was added nitrite and NO. Black line, initial emission spectrum upon addition of 5 μm DAF to the LtHb solution; red line, change in the emission after 20 min; blue line, change in emission after 40 min. B, a control ferrous NO sample in the absence of nitrite as described under “Experimental Procedures.” Black line, initial emission spectrum upon addition of 5 μm DAF; red line, change in the emission after 20 min; blue line, change in emission after 40 min. C, the evolution of the population distribution profile for the sample prepared to promote the formation of the intermediate used with DAF-FM as described under “Experimental Procedures.” D, the comparable evolution of the population distribution profile for the control sample to which DAF-FM was added. inter, intermediate.
Mentions: Figs. 11 and 12 show the evolution of the DAF-FM fluorescence spectra under anaerobic conditions when DAF-FM was added to samples prepared as either the ferrous NO derivative or the NA intermediate of HbA and LtHb. The absorption spectrum was used to establish that in each case the populations were overwhelmingly either ferrous NOHb or the intermediate. For both HbA and LtHb, the increase in fluorescence intensity and the red shift of the fluorescence peak were greater and more rapid for the sample that manifests the absorption spectrum attributable to the NA intermediate (nitrite and NO both added). A similar shift pattern was seen in the absorption maximum of DAF-FM that is also indicative of triazole formation for the samples manifesting the NA intermediate spectrum (not shown).

Bottom Line: A potentially important additional property is the capacity of the HBOC either to generate nitric oxide (NO) or to preserve NO bioactivity to compensate for decreased levels of NO in the circulation.The results show that LtHb undergoes the same reactions as HbA and that the reduced efficacy for these reactions for LtHb relative to HbA is consistent with the much higher redox potential of LtHb.Evidence of functional heterogeneity in LtHb is explained in terms of the large difference in the redox potential of the isolated subunits.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, New York 10461.

Show MeSH
Related in: MedlinePlus