Structural and motional changes in glyceraldehyde-3-phosphate dehydrogenase upon binding to the band-3 protein of the erythrocyte membrane examined with [15N,2H]maleimide spin label and electron paramagnetic resonance.

Beth, Albert H.; Balasubramanian, K.A.; Wilder, Robert T.; Venkataramu, S. D.; Robinson, Bruce H.; Dalton, Larry R.; Pearson, D. E.; Park, Jane H.

doi:10.1073/pnas.78.8.4955

Cited by 28 publications

(5 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Time-resolved phosphorescence experiments using an antibody to the cytoplasmic domain indicated a highly flexible structure in the region of the binding site (McPherson et al, 1992). ST-EPR studies of spin-labeled hemoglobin (Cassoly, 1982) and spin-labeled glyceraldehyde-3-phosphate dehydrogenase (Beth et al, 1981) bound to the N-terminus of the cytoplasmic domain also suggested that there was considerable motional freedom of this region relative to the hindered rotation of the transmembrane domain.…”

Section: Overview Of Previous Workmentioning

confidence: 97%

Flexibility of the Cytoplasmic Domain of the Anion Exchange Protein, Band 3, in Human Erythrocytes

Blackman

Hustedt

Cobb

et al. 2001

Biophysical Journal

View full text Add to dashboard Cite

The rotational flexibility of the cytoplasmic domain of band 3, in the region that is proximal to the inner membrane surface, has been investigated using a combination of time-resolved optical anisotropy (TOA) and saturation-transfer electron paramagnetic resonance (ST-EPR) spectroscopies. TOA studies of rotational diffusion of the transmembrane domain of band 3 show a dramatic decrease in residual anisotropy following cleavage of the link with the cytoplasmic domain by trypsin (E. A. Nigg and R. J. Cherry, 1980, Proc. Natl. Acad. Sci. U.S.A. 77:4702-4706). This result is compatible with two independent hypotheses: 1) trypsin cleavage leads to dissociation of large clusters of band 3 that are immobile on the millisecond time scale, or 2) trypsin cleavage leads to release of a constraint to uniaxial rotational diffusion of the transmembrane domain. ST-EPR studies at X- and Q-band microwave frequencies detect rotational diffusion of the transmembrane domain of band 3 about the membrane normal axis of reasonably large amplitude that does not change upon cleavage with trypsin. These ST-EPR results are not consistent with dissociation of clusters of band 3 as a result of cleavage with trypsin. Global analyses of the ST-EPR data using a newly developed algorithm indicate that any constraint to rotational diffusion of the transmembrane domain of band 3 via interactions of the cytoplasmic domain with the membrane skeleton must be sufficiently weak to allow rotational excursions in excess of 32 degrees full-width for a square-well potential. In support of this result, analyses of the TOA data in terms of restricted amplitude uniaxial rotational diffusion models suggest that the membrane-spanning domain of that population of band 3 that is linked to the membrane skeleton is constrained to diffuse in a square-well of approximately 73 degrees full-width. This degree of flexibility may be necessary for providing the unique mechanical properties of the erythrocyte membrane.

show abstract

Section: Overview Of Previous Workmentioning

confidence: 97%

Flexibility of the Cytoplasmic Domain of the Anion Exchange Protein, Band 3, in Human Erythrocytes

Blackman

Hustedt

Cobb

et al. 2001

Biophysical Journal

View full text Add to dashboard Cite

show abstract

“…The "55 kDa" transmembrane domain (actual molecular weight 59.3 kDa) traverses the bilayer 12 to 14 times thereby forming an anion exchange pathway for chloride and bicarbonate (2). The "41 kDa" cytoplasmic domain of band 3 (cdb3; actual molecular weight 42.5 kDa) serves as an important organizing center for a large number of protein-protein interactions (1,3) including ankyrin (4), protein 4.2 (5), protein 4.1 (6), glycolytic enzymes (7)(8)(9)(10), hemoglobin (11)(12)(13), hemichromes (14), and p72 syk (15). A number of studies carried out over the past two decades have suggested that cdb3 exhibits an elongated, highly asymmetric structure when separated from the transmembrane domain of AE1 as a water soluble protein (16,17) as well as when present in full length AE1 in the erythrocyte membrane (18).…”

Section: Also Known As Band 3 (Reviewed In Ref 1)mentioning

confidence: 99%

Solution Structure of the Cytoplasmic Domain of Erythrocyte Membrane Band 3 Determined by Site-Directed Spin Labeling

et al. 2005

View full text Add to dashboard Cite

The cytoplasmic domain of the anion exchange protein (cdb3) serves as a critical organizing center for protein-protein interactions that stabilize the erythrocyte membrane. The structure of the central core of cdb3, determined by X-ray crystallography from crystals grown at pH 4.8, revealed a compact dimer for residues 55-356 and unresolved N- and C-termini on each monomer [Zhang et al. (2000) Blood 96, 2925-2933]. Given that previous studies had suggested a highly asymmetric structure for cdb3 and that pH dependent structural transitions of cdb3 have been reported, the structure of cdb3 in solution at neutral pH was investigated via site-directed spin labeling in combination with conventional electron paramagnetic resonance (EPR) and double electron electron resonance (DEER) spectroscopies. These studies show that the structure of the central compact dimer (residues 55-356) is indistinguishable from the crystal structure determined at pH 4.8. N-Terminal residues 1-54 and C-terminal residues 357-379 are dynamically disordered and show no indications of stable secondary structure. These results establish a structural model for cdb3 in solution at neutral pH which represents an important next step in characterizing structural details of the protein-protein interactions that stabilize the erythrocyte membrane.

show abstract

“…Istopic substitution of the protons in the spin-label with deuterons reduces inhomogeneous line broadening, increasing the signal-to-noise ratio and the spectral resolution, in particular in the ST-EPR time domain (). Isotopic substitution of 15 N, which has two nuclear spin states, for 14 N, which has three nuclear spin states, simplifies the spectrum, and also increases the signal-to-noise ratio and the spectral resolution ( , ). The earlier EPR study () found that the occupied receptor had a rotational correlation time consistent with smaller oligomers, but was limited in three respects.…”

mentioning

confidence: 99%

Rotational Dynamics of the Epidermal Growth Factor Receptor

2002

View full text Add to dashboard Cite

We have examined the rotational mobility of SL-EGF, a bifunctional adduct of bis(sulfo-N-succinimidyl)-[(15)N,(2)H(16)]-doxyl-2-spiro-4'-pimelate and [Lys3,Tyr22]-murine epidermal growth factor, bound to the EGF receptor in A431 membrane vesicles. The linear EPR spectrum indicated that there was essentially no free SL-EGF in the bound complex preparation. To better define the rotational mobility of the SL-EGF bound to the EGF receptor, ST-EPR spectra were obtained at multiple Zeeman field modulation frequencies. Global analysis with a uniaxial rotational diffusion model of the ST-EPR data yielded two minima that have differences in rotational mobility and in orientation of the SL-EGF relative to the membrane normal axis. The rotational mobilities of the two rotational species are consistent with monomers and dimers or somewhat larger oligomers, such as trimers or tetramers, arguing against a role for higher order receptor clustering in receptor activation. Considering the two minima and previous observations that A431 membrane vesicles contain two distinguishable ligand-binding populations, the ST-EPR spectra were fit with a model having two uniaxial rotating species. This yielded two components that were similar to those obtained from the two original one-component fits, either fast or slow rotational mobility, with different orientations. The model-dependent results obtained suggest that there are potential conformational and rotational differences in the two populations and provide a plausible description for the origin of high- and low-affinity EGF-binding sites that can be tested in future experiments.

show abstract

Structural and motional changes in glyceraldehyde-3-phosphate dehydrogenase upon binding to the band-3 protein of the erythrocyte membrane examined with [15N,2H]maleimide spin label and electron paramagnetic resonance.

Cited by 28 publications

References 16 publications

Flexibility of the Cytoplasmic Domain of the Anion Exchange Protein, Band 3, in Human Erythrocytes

Flexibility of the Cytoplasmic Domain of the Anion Exchange Protein, Band 3, in Human Erythrocytes

Solution Structure of the Cytoplasmic Domain of Erythrocyte Membrane Band 3 Determined by Site-Directed Spin Labeling

Rotational Dynamics of the Epidermal Growth Factor Receptor

Contact Info

Product

Resources

About