A Study of the Mechanisms of Slow Religation to Sickle Cell Hemoglobin Polymers Following Laser Photolysis

Shapiro, Daniel B.; Esquerra, Raymond M.; Goldbeck, Robert A.; Ballas, Samir K.; Mohandas, Narla; Kliger, David S.

doi:10.1006/jmbi.1996.0372

Cited by 7 publications

(10 citation statements)

References 18 publications

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“…Therefore, commercial software, such as SpecWt (Spectrum Software Associates, Marlborough, MA) has been widely applied [41,42,58]. Other studies have been carried out by applying in-house programs [37,40,54,63]. On the other hand, self-modeling can still be considered a tool under development.…”

Section: Softwarementioning

confidence: 98%

“…The pure spectra for both species were recovered by a least squares procedure at each wavelength. SVD has also been applied in hemoglobin studies monitored by time-resolved linear dichroism [40]. A similar procedure was applied for the study of the reaction of hidroxyurea with hemoglobin S monitored by molecular absorption spectroscopy [41].…”

Section: Applications Of Svd and Global Analysis To Biophysical And Bmentioning

confidence: 99%

“…For Hb S, two steps were postulated. SVD and global analysis was applied as in [40,41] and allowed the recovery of the absorption spectrum for the intermediate species.…”

Section: Applications Of Svd and Global Analysis To Biophysical And Bmentioning

confidence: 99%

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Application of multivariate resolution methods to the study of biochemical and biophysical processes

Jaumot

Vives

Gargallo

2004

Analytical Biochemistry

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Section: Softwarementioning

confidence: 98%

Section: Applications Of Svd and Global Analysis To Biophysical And Bmentioning

confidence: 99%

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Application of multivariate resolution methods to the study of biochemical and biophysical processes

Jaumot

Vives

Gargallo

2004

Analytical Biochemistry

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“…However, later studies used the ultrasensitive TRLD technique to study the photocycle of bacteriorhodopsin in purple membrane patches, the results demonstrating that during transition from the K to L intermediate (50 ns to 50 µs) the retinal chromophore rotates less than 8 ° relative to the unphotolyzed chromophore [36, 82], and the rebinding kinetics of sickle hemoglobin polymers, where the rate of photodissociated CO ligand recombination to the polymers was found to be ~1000 times slower than the rate for CO rebinding to T-state hemoglobin monomers [83]. …”

Section: Nanosecond Time-resolved Polarization Spectroscopymentioning

confidence: 99%

Nanosecond time-resolved polarization spectroscopies: Tools for probing protein reaction mechanisms

Chen

Goldbeck

Kliger

2010

Methods

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Polarization methods, introduced in the 1800's, offered one of the earliest ways to examine protein structure. Since then, many other structure-sensitive probes have been developed, but circular dichroism (CD) remains a powerful technique because of its versatility and the specificity of protein structural information that can be explored. With improvements in time-resolution, from millisecond to picosecond CD measurements, it has proven to be an important tool for studying the mechanism of folding and function in many biomolecules. For example, nanosecond time-resolved CD (TRCD) studies of the sub-microsecond events of reduced cytochrome c folding have provided direct experimental evidence of kinetic heterogeneity, which is an inherent property of the diffusional nature of early folding dynamics on the energy landscape. In addition, TRCD has been applied to the study of many biochemical processes, such as ligand rebinding in hemoglobin and myoglobin and signaling state formation in photoactive yellow protein and prototropin 1 LOV2. The basic approach to TRCD has also been extended to include a repertoire of nanosecond polarization spectroscopies: optical rotatory dispersion (ORD), magnetic CD and ORD, and linear dichroism. This article will discuss the details of the polarization methods used in this laboratory, as well as the coupling of timeresolved ORD with the temperature-jump trigger so that protein folding can be studied in a larger number of proteins.

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“…Slow rebinding of the polymer phase has been measured following laser photolysis of a partially CO-saturated gel (Shapiro et al, 1995). Using time-resolved linear dichroism to selectively probe the ligation state of the polymer phase, it was found that the rate of CO rebinding to HbS polymers is ϳ1000 times slower than to solution phase molecules (Shapiro et al, 1995(Shapiro et al, , 1996.…”

Section: Introductionmentioning

confidence: 99%

Sickle Hemoglobin Polymer Melting in High Concentration Phosphate Buffer

Louderback

Ballas

Kim‐Shapiro

1999

Biophysical Journal

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Sickle cell hemoglobin (HbS) prepared in argon-saturated 1.8 M phosphate buffer was rapidly mixed with carbon monoxide (CO)-saturated buffer. The binding of CO to the sickle hemoglobin and the simultaneous melting of the hemoglobin polymers were monitored by transmission spectroscopy (optical absorption and turbidity). Changes in the absorption profile were interpreted as resulting from CO binding to deoxy-HbS while reduced scattering (turbidity) was attributed to melting (depolymerization) of the HbS polymer phase. Analysis of the data provides insight into the mechanism and kinetics of sickle hemoglobin polymer melting. Conversion of normal deoxygenated, adult hemoglobin (HbA) in high concentration phosphate buffer to the HbA-CO adduct was characterized by an average rate of 83 s-1. Under the same conditions, conversion of deoxy-HbS in the polymer phase to the HbS-CO adduct in the solution phase is characterized by an average rate of 5.8 s-1 via an intermediate species that grows in with a 36 s-1 rate. Spectral analysis of the intermediate species suggests that a significant amount of CO may bind to the polymer phase before the polymer melts.

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A Study of the Mechanisms of Slow Religation to Sickle Cell Hemoglobin Polymers Following Laser Photolysis

Cited by 7 publications

References 18 publications

Application of multivariate resolution methods to the study of biochemical and biophysical processes

Application of multivariate resolution methods to the study of biochemical and biophysical processes

Nanosecond time-resolved polarization spectroscopies: Tools for probing protein reaction mechanisms

Sickle Hemoglobin Polymer Melting in High Concentration Phosphate Buffer

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