Structural stability of the erythrocyte anion transporter, band 3, in native membranes and in detergent micelles

Sami, Malkit; Malik, Saira; Watts, Anthony

doi:10.1016/0005-2736(92)90173-j

Cited by 24 publications

(5 citation statements)

References 27 publications

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“…A lower value of 1.3 cal/g was found for the denaturation enthalpy of Na + ,K + -ATPase (16). Higher enthalpy values for the band 3 protein from erythrocytes had been reported; however, it has been found that if this protein is stripped of peripheral membrane proteins, the observed enthalpy of denaturation is 3.5 cal/g (26,27), in line with values for other integral membrane proteins. The value of about 3 cal/g for integral membrane proteins compares with the much larger enthalpy for the denaturation of globular proteins at 67 °C, which is 7.8 ( 0.7 cal/g (28).…”

Section: Discussionsupporting

confidence: 65%

Glucose-Induced Thermal Stabilization of the Native Conformation of GLUT 1

Epand

Jung

1998

Biochemistry

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The glucose transporter, GLUT 1, was purified from erythrocyte membranes and incorporated into vesicles of erythrocyte lipids. These protein-containing vesicles were studied with differential scanning calorimetry. It was found that the protein underwent an irreversible denaturation at 68.5 +/- 0.2 degreesC (at a scan rate of 0.25 degreesC/min) which was shifted to 72.6 +/- 0.2 degreesC in the presence of 500 mM D-glucose, while 500 mM L-glucose or 10 microM cytochalasin B did not produce a significant shift. The calorimetric enthalpy was found to be 150 kcal/mol, independent of the presence of D-glucose. On a weight basis this value is lower than that for soluble proteins, but it is comparable to values obtained with other integral membrane proteins. The van't Hoff enthalpy is similar to the calorimetric enthalpy, within the experimental error, indicating that the transition is not likely to be cooperative. The activation energy is estimated from both the scan rate dependence of the transition temperature and from the shape of the DSC curve. The presence of 500 mM D-glucose slightly decreases the activation energy. It is concluded that the shift to a higher denaturation transition temperature in the presence of D-glucose is not a result of increased kinetic stability of GLUT 1.

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Section: Discussionsupporting

confidence: 65%

Glucose-Induced Thermal Stabilization of the Native Conformation of GLUT 1

Epand

Jung

1998

Biochemistry

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“…Finally, the choice of detergent may be important in stabilizing a particular oligomeric form of the protein. Calorimetric analysis of band 3 in both TX-100 and C 12 E 8 indicated a similar stabilization of the protein in the two detergents (Sami et al, 1992). However, the stability of different oligomeric forms of the protein in the different detergents was not examined.…”

Section: Discussionmentioning

confidence: 94%

Hydrodynamic Properties of Human Erythrocyte Band 3 Solubilized in Reduced Triton X-100

Taylor

Boulter

Harding

et al. 1999

Biophysical Journal

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The oligomeric state and function of band 3, purified by sulfhydryl affinity chromatography in reduced Triton X-100, was investigated. Size exclusion high-performance liquid chromatography showed that a homogeneous population of band 3 dimers could be purified from whole erythrocyte membranes. The elution profile of band 3 purified from membranes that had been stripped of its cytoskeleton before solubilization was a broad single peak describing a heterogeneous population of oligomers with a mean Stokes radius of 100 A. Sedimentation velocity ultracentrifugation analysis confirmed particle heterogeneity and further showed monomer/dimer/tetramer equilibrium self-association. Whether the conversion of dimer to the form described by a Stokes radius of 100 A was initiated by removal of cytoskeletal components, alkali-induced changes in band 3 conformation, or alkali-induced loss of copurifying ligands remains unclear. After incubation at 20 degrees C for 24 h, both preparations of band 3 converted to a common form characterized by a mean Stokes radius of 114 A. This form of the protein, examined by equilibrium sedimentation ultracentrifugation, is able to self-associate reversibly, and the self-association can be described by a dimer/tetramer/hexamer model, although the presence of higher oligomers cannot be discounted. The ability of the different forms of the protein to bind stilbene disulfonates revealed that the dimer had the highest inhibitor binding affinity, and the form characterized by a mean Stokes radius of 114 A to have the lowest.

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“…Therefore, if detergents interact with band 3 in a way similar to fatty acids, then a detergent with a long alkyl chain might provide an excellent environment for band 3. However, recent evidence from calorimetry studies of band 3 in polyoxyethylene detergents with chain lengths Cio-Cig shows that Ci 2 provides the optimal alkyl chain length for thermal stability of band 3 (Sami et al, 1992).…”

Section: Discussionmentioning

confidence: 99%

Detergent interaction with band 3, a model polytopic membrane protein

Casey¹,

Reithmeier²

1993

Biochemistry

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The interaction of band 3, the 95-kDa anion-exchange protein of the human erythrocyte membrane, with a variety of nonionic detergents was studied. Band 3 dimers (Stokes radius = 76 A) prepared in octaethylene glycol monododecyl ether (C12E8) could be exchanged into a variety of detergents by size-exclusion high-performance liquid chromatography (HPLC), with complete removal of C12E8 from band 3 being confirmed using radiolabeled detergent. Critical micellar concentration (cmc) values, determined for all detergents in the buffer used for HPLC analysis, ranged from 0.47 microM to 223 mM. Band 3 was found to aggregate in all detergents below their cmc, and concentrations of detergents 2-200 times the cmc were required to prevent aggregation. For detergents with a low cmc, it was important to ensure that the concentration of detergent micelles minimally equalled the concentration of protein. Hydrodynamic measurements and cross-linking studies showed that band 3 remained dimeric in most detergents above their cmc. Furthermore, circular dichroism and inhibitor binding studies supported the view that band 3 can retain its native structure after detergent exchange. Detergents with short alkyl chains (C8) denature band 3, while detergents with longer alkyl chains (C12) maintained the native structure of band 3. The ability to exchange band 3 into a variety of detergents with the maintenance of native structure is an essential prerequisite for crystallization trials. The results obtained in this study of band 3, a model polytopic (multispanning) membrane protein, may be generally applicable to other membrane proteins.

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Structural stability of the erythrocyte anion transporter, band 3, in native membranes and in detergent micelles

Cited by 24 publications

References 27 publications

Glucose-Induced Thermal Stabilization of the Native Conformation of GLUT 1

Glucose-Induced Thermal Stabilization of the Native Conformation of GLUT 1

Hydrodynamic Properties of Human Erythrocyte Band 3 Solubilized in Reduced Triton X-100

Detergent interaction with band 3, a model polytopic membrane protein

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