Direct intracellular measurement of deoxygenated hemoglobin S solubility

Fabry, Mary E.; Desrosiers, Laurephile; Suzuka, Sandra M.

doi:10.1182/blood.v98.3.883

Cited by 19 publications

(12 citation statements)

References 6 publications

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“…Our recent observations, combined with earlier reports (Vedvick et al, 1975;Poillon and Bertles, 1977;Roth et al, 1979;Bookchin et al, 1999;Yohe et al, 2000;Fabry et al, 2001), suggest that certain precautions be taken when using the high phosphate model. Although in our system, we did not observe increases in turbidity due to protein aggregation, these aggregates may produce measurable turbidity in some spectrometers or conditions since the measured turbidity depends on the solid angle measured by the detector and on the intensity and angular dependence of scattered light from the aggregates.…”

Section: Discussionsupporting

confidence: 74%

“…Although many similarities between HbS polymerization in the high phosphate model system and that under more physiologically relevant conditions have been demonstrated, some cautionary notes related to the use of the high phosphate model have also been made (Vedvick et al, 1975;Poillon and Bertles, 1977;Roth et al, 1979;Bookchin et al, 1999;Fabry et al, 2001). These studies demonstrate that, although the high phosphate model provides a convenient way to assess affects on polymerization, caution is needed when applying results from its use to physiology.…”

Section: Introductionmentioning

confidence: 99%

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Aggregation of Normal and Sickle Hemoglobin in High Concentration Phosphate Buffer

Chen

Ballas

Hantgan

et al. 2004

Biophysical Journal

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Sickle cell disease is caused by a mutant form of hemoglobin, hemoglobin S, that polymerizes under hypoxic conditions. The extent and mechanism of polymerization are thus the subject of many studies of the pathophysiology of the disease and potential treatment strategies. To facilitate such studies, a model system using high concentration phosphate buffer (1.5 M-1.8 M) has been developed. To properly interpret results from studies using this model it is important to understand the similarities and differences in hemoglobin S polymerization in the model compared to polymerization under physiological conditions. In this article, we show that hemoglobin S and normal adult hemoglobin, hemoglobin A, aggregate in high concentration phosphate buffer even when the concentration of hemoglobin is below the solubility defined for polymerization. This phenomenon was not observed using 0.05 M phosphate buffer or in another model system we studied that uses dextran to enhance polymerization. We have used static light scattering, dynamic light scattering, and differential interference contrast microscopy to confirm aggregation of deoxygenated and oxygenated hemoglobins below their solubility and have shown that this aggregation is not observable using turbidity measurements, a common technique for assessing polymerization. We have also shown that the aggregation increases with increasing temperature in the range of 15 degrees -37 degrees C and that it increases as the concentration of phosphate increases. These studies contribute to the working knowledge of how to properly apply studies of hemoglobin S polymerization that are conducted using the high phosphate model.

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Section: Discussionsupporting

confidence: 74%

Section: Introductionmentioning

confidence: 99%

Aggregation of Normal and Sickle Hemoglobin in High Concentration Phosphate Buffer

Chen

Ballas

Hantgan

et al. 2004

Biophysical Journal

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“…One possible interpretation of these and our findings is that increased cerebral blood flow (CBF) and decreased concentration of deoxyhemoglobin in SCA might have been at the base of the diminished SWI contrast and venular conspicuity in SCA patients. Prior research, however, shows that although the oxygen affinity of haemoglobin S is decreased in its polymerised state and in hypoxic conditions, it is near normal in its tetrameric state and in patients in normoxia (Abdu et al, 2008;Fabry et al, 2001), like the steady state SCA patients in our study. In children, lower peripheral oxygen saturation was positively correlated with cerebral desaturation (Quinn and Dowling, 2012).…”

Section: Discussioncontrasting

confidence: 41%

Brain venular pattern by 7T MRI correlates with memory and haemoglobin in sickle cell anaemia

Novelli

Sarles

Aizenstein

et al. 2015

Psychiatry Research: Neuroimaging

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Sickle cell anaemia (SCA) is a hereditary hemoglobinopathy characterised by extensive vascular dysfunction that stems from inflammation, thrombosis and occlusion of post-capillary venules. Cognitive impairment is a neurological complication of SCA whose pathogenesis is unknown. We hypothesised that cerebral venular abnormalities are linked to cognitive impairment in SCA. Thus, we employed 7T magnetic resonance imaging (MRI) to examine the association between venular density and cognitive function in homozygous SCA. We quantified the density of total, long, and short venules in pre-defined regions of interest between the frontal and occipital cornu on each hemisphere. Cognitive function was assessed using the Hopkins Verbal Learning Test - Revised (HVLT-R) test of learning and memory. Patients (n=11) were compared with race, age and gender-equated controls (n=7). Compared to controls, patients had an overall venular rarefaction, with significantly lower density of long venules and greater density of short venules which was inversely related to HVLT-R performance and haemoglobin. To our knowledge, this is the first 7T MRI study in SCA and first report of associations between cerebral venular patterns and cognitive performance and haemoglobin. Future studies should examine whether these novel neuroimaging markers predict cognitive impairment longitudinally and are mechanistically linked to severity of anaemia.

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“…The solubility of deoxy-Hb S, which is the concentration of fully deoxy-Hb S in equilibrium with polymer (C SAT ), is a factor that determines polymer formation in vivo (Fabry et al, 2001(Fabry et al, , 2003. Measurement of C SAT with INN-312 showed a significant elevation in the solubility of deoxy-Hb S, which increased from 0.015 g dl À1 (0 mM INN-312), to 0.017, 0.025 and >0.05 g dl À1 in samples incubated with 0.5, 1 or 2 mM INN-312, respectively (Fig.…”

Section: Inn Compounds Show Highly Potent And/or Dual Antisickling Acmentioning

confidence: 99%

Crystallographic analysis of human hemoglobin elucidates the structural basis of the potent and dual antisickling activity of pyridyl derivatives of vanillin

Abdulmalik

Ghatge

Musayev

et al. 2011

Acta Crystallogr D Biol Cryst

135

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Vanillin has previously been studied clinically as an antisickling agent to treat sickle-cell disease. In vitro investigations with pyridyl derivatives of vanillin, including INN-312 and INN-298, showed as much as a 90-fold increase in antisickling activity compared with vanillin. The compounds preferentially bind to and modify sickle hemoglobin (Hb S) to increase the affinity of Hb for oxygen. INN-312 also led to a considerable increase in the solubility of deoxygenated Hb S under completely deoxygenated conditions. Crystallographic studies of normal human Hb with INN-312 and INN-298 showed that the compounds form Schiff-base adducts with the N-terminus of the α-subunits to constrain the liganded (or relaxed-state) Hb conformation relative to the unliganded (or tense-state) Hb conformation. Interestingly, while INN-298 binds and directs its meta-positioned pyridine-methoxy moiety (relative to the aldehyde moiety) further down the central water cavity of the protein, that of INN-312, which is ortho to the aldehyde, extends towards the surface of the protein. These studies suggest that these compounds may act to prevent sickling of SS cells by increasing the fraction of the soluble high-affinity Hb S and/or by stereospecific inhibition of deoxygenated Hb S polymerization.

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Direct intracellular measurement of deoxygenated hemoglobin S solubility

Cited by 19 publications

References 6 publications

Aggregation of Normal and Sickle Hemoglobin in High Concentration Phosphate Buffer

Aggregation of Normal and Sickle Hemoglobin in High Concentration Phosphate Buffer

Brain venular pattern by 7T MRI correlates with memory and haemoglobin in sickle cell anaemia

Crystallographic analysis of human hemoglobin elucidates the structural basis of the potent and dual antisickling activity of pyridyl derivatives of vanillin

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