Imaging of the diffusion of single band 3 molecules on normal and mutant erythrocytes

Kodippili, Gayani C.; Spector, Jeff; Sullivan, Caitlin; Kuypers, Frans A.; Labotka, Richard; Gallagher, Patrick G.; Ritchie, Ken; Low, Philip S.

doi:10.1182/blood-2009-02-205450

Cited by 77 publications

(125 citation statements)

References 46 publications

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“…Population Analysis of SPT Data-We and others have used SPT on lymphocytes (18,19) and erythrocytes (29,30) and have found heterogeneous distributions of diffusion coefficients for membrane proteins. Accordingly, data from these experiments should be analyzed for the appearance of subpopulations that may reveal additional information about receptor mobility.…”

Section: Resultsmentioning

confidence: 99%

Dynamic Regulation of CD45 Lateral Mobility by the Spectrin-Ankyrin Cytoskeleton of T Cells

Cairo

Das

Albohy

et al. 2010

Journal of Biological Chemistry

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The leukocyte common antigen, CD45, is a critical immune regulator whose activity is modulated by cytoskeletal interactions. Components of the spectrin-ankyrin cytoskeleton have been implicated in the trafficking and signaling of CD45. We have examined the lateral mobility of CD45 in resting and activated T lymphocytes using single-particle tracking and found that the receptor has decreased mobility caused by increased cytoskeletal contacts in activated cells. Experiments with cells that have disrupted ␤⌱ spectrin interactions show decreased cytoskeletal contacts in resting cells and attenuation of receptor immobilization in activated cells. Applying two types of population analyses to single-particle tracking trajectories, we find good agreement between the diffusion coefficients obtained using either a mean squared displacement analysis or a hidden Markov model analysis. Hidden Markov model analysis also reveals the rate of association and dissociation of CD45-cytoskeleton contacts, demonstrating the importance of this analysis for measuring cytoskeleton binding events in live cells. Our findings are consistent with a model in which multiple cytoskeletal contacts, including those with spectrin and ankyrin, participate in the regulation of CD45 lateral mobility. These interactions are a major factor in CD45 immobilization in activated cells. Furthermore, cellular activation leads to CD45 immobilization by reduction of the CD45-cytoskeleton dissociation rate. Short peptides that mimic spectrin repeat domains alter the association rate of CD45 to the cytoskeleton and cause an apparent decrease in dissociation rates. We propose a model for CD45-cytoskeleton interactions and conclude that the spectrin-ankyrin-actin network is an essential determinant of immunoreceptor mobility.The lymphocyte receptor-like protein tyrosine phosphatase, CD45, 2 is a transmembrane protein that acts as a critical component of T cell activation (1). The phosphatase activity of the CD45 cytoplasmic domain contributes to T cell activation through the removal of an inhibitory phosphate group on a membrane-associated Src family kinase, such as Tyr 505 of p56 lck (Lck) (2). Other CD45 substrates have been established in addition to Lck, including Fyn, SKAP-55, JAK, ZAP-70, and CK2 (1). The involvement of CD45 as a regulator of critical signaling pathways and autoimmune diseases highlights the importance of understanding its function.In addition to phosphoprotein substrates, CD45 interacts with a variety of molecular partners that modulate its activity. Its heavily glycosylated extracellular domain interacts with Thy1 (3) and galectins (4), and it self-associates to form dimers. Dimer formation is a process linked to both regulation of its phosphatase activity and sialylation of its ectodomain (1, 5, 6). The cytoplasmic domain of CD45 contains one catalytically active protein-tyrosine phosphatase domain (Domain 1) and one inactive domain (Domain 2) and binds to key cellular regulators of immune function such as CD45AP (7), CD2 (8), CD11a (9),...

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Section: Resultsmentioning

confidence: 99%

Dynamic Regulation of CD45 Lateral Mobility by the Spectrin-Ankyrin Cytoskeleton of T Cells

Cairo

Das

Albohy

et al. 2010

Journal of Biological Chemistry

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“…32 These incubation conditions allowed for covalent reaction of the compound with no more than 2 band 3 molecules per cell. DIDS-biotin-labeled cells were incubated with excess streptavidinconjugated Qdot525 (Invitrogen) and washed 2 times with PBS containing 0.1% BSA.…”

Section: Labeling Of Band 3 In Whole Erythrocytes With Dids-biotin Fomentioning

confidence: 99%

“…Video recordings of individual band 3 molecules were collected for 1000 frames at 120 frames/s, and analyzed essentially as described previously. 32 Band 3 diffusion was recorded from 0-60 minutes after the addition of o-vanadate, but only data from 30-60 minutes of treatment were ultimately used because phosphorylation was found to be maximal during this time period.…”

Section: Labeling Of Band 3 In Whole Erythrocytes With Dids-biotin Fomentioning

confidence: 99%

“…To confirm this hypothesis, we measured the diffusion of band 3 in untreated and o-vanadate-treated erythrocytes using single-particle tracking methods. 32 Two diffusion coefficients were measured, one corresponding to band 3 diffusion over short time intervals, termed the microscopic diffusion coefficient (D micro ), and the other corresponding to its asymptotic diffusion coefficient at long time points, termed the macroscopic diffusion coefficient (D Macro ). The first, D micro , provides information on the more rapid motions of the protein, which could include simple Brownian motion or oscillation of band 3 while attached to a flexible tether.…”

Section: Band 3 Diffusion In Intact O-vanadate-treated Erythrocytesmentioning

confidence: 99%

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Regulation of membrane-cytoskeletal interactions by tyrosine phosphorylation of erythrocyte band 3

Ferru

Giger²,

Pantaleo

et al. 2011

Blood

Self Cite

172

216

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The cytoplasmic domain of band 3 serves as a center of erythrocyte membrane organization and constitutes the major substrate of erythrocyte tyrosine kinases. Tyrosine phosphorylation of band 3 is induced by several physiologic stimuli, including malaria parasite invasion, cell shrinkage, normal cell aging, and oxidant stress (thalassemias, sickle cell disease, glucose-6-phosphate dehydrogenase deficiency, etc). In an effort to characterize the biologic sequelae of band 3 tyrosine phosphorylation, we looked for changes in the polypeptide's function that accompany its phosphorylation. We report that tyrosine phosphorylation promotes dissociation of band 3 from the spectrin-actin skeleton as evidenced by: (1) a decrease in ankyrin affinity in direct binding studies, (2) an increase in detergent extractability of band 3 from ghosts, (3) a rise in band 3 cross-linkability by bis-sulfosuccinimidylsuberate, (4) significant changes in erythrocyte morphology, and (5) elevation of the rate of band 3 diffusion in intact cells. Because release of band 3 from its ankyrin and adducin linkages to the cytoskeleton can facilitate changes in multiple membrane properties, tyrosine phosphorylation of band 3 is argued to enable adaptive changes in erythrocyte biology that permit the cell to respond to the above stresses. (Blood. 2011; 117(22):5998-6006) IntroductionEarly views of the human erythrocyte argued that the cell was inert to external stimuli and that its complement of protein kinases, phospholipases, G proteins, phosphatases, and hormone receptors simply constituted nonfunctional vestiges of signaling pathways that were once operational in erythroid precursor cells. More recent evidence, however, has revealed that the human erythrocyte is highly responsive to its environment and that the cell's rich ensemble of signaling proteins likely comprise critical components in the cell's communication with its extracellular milieu. Classic hormones/signaling molecules such as prostaglandin E 2 , insulin, epinephrine, endothelin, ADP, and NO are now known to modulate erythrocyte properties in an adaptive manner, and the functional activities of many intracellular signaling intermediates have been demonstrated to regulate erythrocyte behavior. [1][2][3][4][5] One of the major targets of erythrocyte signaling appears to be the predominant membrane-spanning protein, band 3. Band 3 (AE1) catalyzes the exchange of anions (primarily HCO 3 Ϫ for Cl Ϫ ) across the erythrocyte membrane, 6 anchors the spectrin/ actin cytoskeleton to the lipid bilayer, 7 organizes and regulates a complex of glycolytic enzymes, 8,9 participates in control of erythrocyte lifespan, 10,11 nucleates several important membrane-spanning proteins, 12 and serves as a docking site for multiple peripheral membrane proteins, including protein 4.1, protein 4.2, and several kinases and phosphatases. [13][14][15][16] Not surprisingly, mutations in band 3 are frequently associated with various hemolytic diseases. 17 Perhaps because of its many important functions, band 3 is a...

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“…Since the lipid composition of altered RBC is normal, the acanthocytic shape change results from impaired interaction of the membrane multiprotein complexes (MMPC) with the cytoskeleton of red blood cell 33 . The major RBC anion exchanger protein, band 3 (B3), one of the most abundant membrane protein, is organized either as a tetrameric B3-ankyrin complex, a dimeric B3-protein 4.1R complex (also called "junctional complex") and as a free B3 protein 34,35 . The …”

Section: Distinct Morphology Of Mcleod Erythrocytesmentioning

confidence: 99%

Neurodegeneration in the elderly – When the blood type matters: An overview of the McLeod syndrome with focus on hematological features

Frey

Gassner

Jung

2015

Transfusion and Apheresis Science

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Multisystem deterioration occurs mainly in older individuals and may be related to physiological tissue degeneration. However, genetic predisposition may be unmasked by inappropriate functional and structural system deficiencies. McLeod syndrome (MLS) is a rare, multisystem disease which is X-chromosomal inherited and belongs to the neuroacanthocytosis syndromes (NAS). The main clinical manifestations contain progressive neuro-psychiatric and cognitive deterioration, choreatic movement disorder, as well as myopathy, sensory motor axonal neuropathy and cardiomyopathy. In addition, MLS patients have red blood cell abnormalities including immune-hematological, morphological and functional impairments of red blood cells. In large deletions, contiguous gene syndrome may arise, including Duchenne muscular dystrophia, cellular immunodeficiency or retinitis pigmentosa. Hematological abnormalities such as blood group abnormalities in Kell-and XK blood group system, formation of anti-public red blood cell alloantibodies, acanthocytosis and elevated creatinine phosphokinase may precede clinical disease manifestation for decades and provide tools for early diagnosis. Patients with unexplained neuro-muscular deterioration and/or neuro-psychological pathologies accompanied with hematological abnormalities should be investigated for MLS.

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Imaging of the diffusion of single band 3 molecules on normal and mutant erythrocytes

Cited by 77 publications

References 46 publications

Dynamic Regulation of CD45 Lateral Mobility by the Spectrin-Ankyrin Cytoskeleton of T Cells

Dynamic Regulation of CD45 Lateral Mobility by the Spectrin-Ankyrin Cytoskeleton of T Cells

Regulation of membrane-cytoskeletal interactions by tyrosine phosphorylation of erythrocyte band 3

Neurodegeneration in the elderly – When the blood type matters: An overview of the McLeod syndrome with focus on hematological features

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