scholarly journals Band 3 Erythrocyte Membrane Protein Acts as Redox Stress Sensor Leading to Its Phosphorylation by p72Syk

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Antonella Pantaleo ◽  
Emanuela Ferru ◽  
Maria Carmina Pau ◽  
Amina Khadjavi ◽  
Giorgia Mandili ◽  
...  
Keyword(s):  
Cytoplasmic Domain ◽  
Temporal Variations ◽  
Band 3 ◽  
Serine Phosphorylation ◽  
Redox Stress ◽  
Redox Sensor ◽  
Syk Kinase ◽  
Irreversible Oxidation ◽  
Syk Inhibitor

In erythrocytes, the regulation of the redox sensitive Tyr phosphorylation of band 3 and its functions are still partially defined. A role of band 3 oxidation in regulating its own phosphorylation has been previously suggested. The current study provides evidences to support this hypothesis: (i) in intact erythrocytes, at 2 mM concentration of GSH, band 3 oxidation, and phosphorylation, Syk translocation to the membrane and Syk phosphorylation responded to the same micromolar concentrations of oxidants showing identical temporal variations; (ii) the Cys residues located in the band 3 cytoplasmic domain are 20-fold more reactive than GSH; (iii) disulfide linked band 3 cytoplasmic domain docks Syk kinase; (iv) protein Tyr phosphatases are poorly inhibited at oxidant concentrations leading to massive band 3 oxidation and phosphorylation. We also observed that hemichromes binding to band 3 determined its irreversible oxidation and phosphorylation, progressive hemolysis, and serine hyperphosphorylation of different cytoskeleton proteins. Syk inhibitor suppressed the phosphorylation of band 3 also preventing serine phosphorylation changes and hemolysis. Our data suggest that band 3 acts as redox sensor regulating its own phosphorylation and that hemichromes leading to the protracted phosphorylation of band 3 may trigger a cascade of events finally leading to hemolysis.

scholarly journals Role of Cytoplasmic Domain Serines in Intracellular Trafficking of Furin

2004 ◽  
Vol 15 (6) ◽  
pp. 2884-2894 ◽  
Author(s):  
Florencia B. Schapiro ◽  
Thwe Thwe Soe ◽  
William G. Mallet ◽  
Frederick R. Maxfield
Keyword(s):  
Plasma Membrane ◽  
Steady State ◽  
Transmembrane Protein ◽  
Interleukin 2 ◽  
Chimeric Protein ◽  
Cytoplasmic Domain ◽  
Serine Phosphorylation ◽  
Endocytic Recycling ◽  
Late Endosomes

Furin is a transmembrane protein that cycles between the plasma membrane, endosomes, and the trans-Golgi network, maintaining a predominant distribution in the latter. It has been shown previously that Tac-furin, a chimeric protein expressing the extracellular and transmembrane domains of the interleukin-2 receptor α chain (Tac) and the cytoplasmic domain of furin, is delivered from the plasma membrane to the TGN through late endosomes, bypassing the endocytic recycling compartment. Tac-furin also recycles in a loop between the TGN and late endosomes. Localization of furin to the TGN is modulated by a six-amino acid acidic cluster that contains two phosphorylatable serines (SDSEED). We investigated the role of these serines in the trafficking of Tac-furin by using a mutant chimera in which the SDS sequence was replaced by the nonphosphorylatable sequence ADA (Tac-furin/ADA). Although the mutant construct is internalized and delivered to the TGN, both the postendocytic trafficking and the steady-state distribution were found to differ from the wild-type. In contrast with Tac-furin, Tac-furin/ADA does not enter late endosomes after being internalized. Instead, it traffics with transferrin to the endocytic recycling compartment, and from there it is delivered to the TGN. As with Tac-furin, Tac-furin/ADA is sorted from the TGN into late endosomes at steady state, but its retrieval from the late endosomes to the TGN is inhibited. These results suggest that serine phosphorylation plays an important role in at least two steps of Tac-furin trafficking, acting as an active sorting signal that mediates the selective sorting of Tac-furin into late endosomes after internalization, as well as its retrieval from late endosomes back to the TGN.

scholarly journals Regulation of alpha 6 beta 1 integrin laminin receptor function by the cytoplasmic domain of the alpha 6 subunit.

1993 ◽  
Vol 123 (4) ◽  
pp. 1017-1025 ◽  
Author(s):  
L M Shaw ◽  
A M Mercurio
Keyword(s):  
Point Mutations ◽  
Surface Expression ◽  
Cytoplasmic Domain ◽  
Serine Phosphorylation ◽  
Laminin Receptor ◽  
Site Directed Mutagenesis ◽  
Receptor Function ◽  
Serine Residue ◽  
Beta 1 Integrin

The alpha 6 beta 1 integrin is expressed on the macrophage surface in an inactive state and requires cellular activation with PMA or cytokines to function as a laminin receptor (Shaw, L. M., J. M. Messier, and A. M. Mercurio. 1990. J. Cell Biol. 110:2167-2174). In the present study, the role of the alpha 6 subunit cytoplasmic domain in alpha 6 beta 1 integrin activation was examined. The use of P388D1 cells, an alpha 6-integrin deficient macrophage cell line, facilitated this analysis because expression of either the alpha 6A or alpha 6B subunit cDNAs restores their activation responsive laminin adhesion (Shaw, L. S., M. Lotz, and A. M. Mercurio. 1993. J. Biol. Chem. 268:11401-11408). A truncated alpha 6 cDNA, alpha 6-delta CYT, was constructed in which the human cytoplasmic domain sequence was deleted after the GFFKR pentapeptide. Expression of this cDNA in P388D1 cells resulted in the surface expression of a chimeric alpha 6-delta CYT beta 1 integrin that was unable to mediate laminin adhesion or increase this adhesion in response to PMA under normal conditions, i.e., in medium that contained physiological concentrations of Ca++ and Mg++. The alpha 6A-delta CYT transfectants adhered to laminin, however, when Ca++/Mg++ was replaced with 150 microM Mn++. We also assessed the role of serine phosphorylation in the regulation of alpha 6A beta 1 integrin function by site-directed mutagenesis of the two serine residues present in the alpha 6A cytoplasmic domain because this domain is phosphorylated on serine residues in response to stimuli that activate the laminin receptor function of alpha 6 A beta 1. Point mutations were introduced in the alpha 6A cDNA that changed either serine residue #1064 (M1) or serine residue #1071 (M2) to alanine residues. In addition, a double mutant (M3) was constructed in which both serine residues were changed to alanine residues. P388D1 transfectants which expressed these serine mutations adhered to laminin in response to PMA to the same extent as cells transfected with wild-type alpha 6A cDNA. These findings provide evidence for a novel mode of integrin regulation that is distinct from that reported for other regulated integrins (O'Toole, T. E., D. Mandelman, J. Forsyth, S. J. Shattil, E. F. Plow, and M. H. Ginsberg. 1991. Science (Wash. DC). 254:845-847. Hibbs, M. L., H. Xu, S. A. Stacker, and T. A. Springer. 1991. Science (Wash. DC). 251:1611-1613), and they demonstrate that serine phosphorylation of the alpha 6A cytoplasmic domain is not involved in this regulation.

scholarly journals Syndecan Transmembrane Domain Specifically Regulates Downstream Signaling Events of the Transmembrane Receptor Cytoplasmic Domain

2021 ◽  
Vol 22 (15) ◽  
pp. 7918
Author(s):  
Jisun Hwang ◽  
Bohee Jang ◽  
Ayoung Kim ◽  
Yejin Lee ◽  
Joonha Lee ◽  
...  
Keyword(s):  
Transmembrane Domain ◽  
Growth Factor Receptor ◽  
Cytoplasmic Domain ◽  
Nih3t3 Cells ◽  
C Elegans ◽  
Downstream Signaling ◽  
Downstream Effectors ◽  
Signaling Events

Despite the known importance of the transmembrane domain (TMD) of syndecan receptors in cell adhesion and signaling, the molecular basis for syndecan TMD function remains unknown. Using in vivo invertebrate models, we found that mammalian syndecan-2 rescued both the guidance defects in C. elegans hermaphrodite-specific neurons and the impaired development of the midline axons of Drosophila caused by the loss of endogenous syndecan. These compensatory effects, however, were reduced significantly when syndecan-2 dimerization-defective TMD mutants were introduced. To further investigate the role of the TMD, we generated a chimera, 2eTPC, comprising the TMD of syndecan-2 linked to the cytoplasmic domain of platelet-derived growth factor receptor (PDGFR). This chimera exhibited SDS-resistant dimer formation that was lost in the corresponding dimerization-defective syndecan-2 TMD mutant, 2eT(GL)PC. Moreover, 2eTPC specifically enhanced Tyr 579 and Tyr 857 phosphorylation in the PDGFR cytoplasmic domain, while the TMD mutant failed to support such phosphorylation. Finally, 2eTPC, but not 2eT(GL)PC, induced phosphorylation of Src and PI3 kinase (known downstream effectors of Tyr 579 phosphorylation) and promoted Src-mediated migration of NIH3T3 cells. Taken together, these data suggest that the TMD of a syndecan-2 specifically regulates receptor cytoplasmic domain function and subsequent downstream signaling events controlling cell behavior.

scholarly journals CO2 dynamics and heterogeneity in a cave atmosphere: role of ventilation patterns and airflow pathways

2021 ◽  
Author(s):  
Lovel Kukuljan ◽  
Franci Gabrovšek ◽  
Matthew D. Covington ◽  
Vanessa E. Johnston
Keyword(s):  
Temporal Variations ◽  
Observation Point ◽  
Spatially Distributed ◽  
Combined Action ◽  
Karst Systems ◽  
The Relationship ◽  
Co2 Dynamics ◽  
Global Carbon

AbstractUnderstanding the dynamics and distribution of CO2 in the subsurface atmosphere of carbonate karst massifs provides important insights into dissolution and precipitation processes, the role of karst systems in the global carbon cycle, and the use of speleothems for paleoclimate reconstructions. We discuss long-term microclimatic observations in a passage of Postojna Cave, Slovenia, focusing on high spatial and temporal variations of pCO2. We show (1) that the airflow through the massif is determined by the combined action of the chimney effect and external winds and (2) that the relationship between the direction of the airflow, the geometry of the airflow pathways, and the position of the observation point explains the observed variations of pCO2. Namely, in the terminal chamber of the passage, the pCO2 is low and uniform during updraft, when outside air flows to the site through a system of large open galleries. When the airflow reverses direction to downdraft, the chamber is fed by inlets with diverse flow rates and pCO2, which enter via small conduits and fractures embedded in a CO2-rich vadose zone. If the spatial distribution of inlets and outlets produces minimal mixing between low and high pCO2 inflows, high and persistent gradients in pCO2 are formed. Such is the case in the chamber, where vertical gradients of up to 1000 ppm/m are observed during downdraft. The results presented in this work provide new insights into the dynamics and composition of the subsurface atmosphere and demonstrate the importance of long-term and spatially distributed observations.

scholarly journals Cooperative Role of the Membrane-proximal and -distal Residues of the Integrin β3 Cytoplasmic Domain in Regulation of Talin-mediated αIIbβ3 Activation

2008 ◽  
Vol 283 (9) ◽  
pp. 5662-5668 ◽  
Author(s):  
Takaaki Hato ◽  
Jun Yamanouchi ◽  
Tatsushiro Tamura ◽  
Yoshihiro Yakushijin ◽  
Ikuya Sakai ◽  
...  
Keyword(s):  
Cytoplasmic Domain ◽  
Integrin Β3

scholarly journals Syk inhibitors interfere with erythrocyte membrane modification during P falciparum growth and suppress parasite egress

Blood ◽  
2017 ◽  
Vol 130 (8) ◽  
pp. 1031-1040 ◽  
Author(s):  
Antonella Pantaleo ◽  
Kristina R. Kesely ◽  
Maria Carmina Pau ◽  
Ioannis Tsamesidis ◽  
Evelin Schwarzer ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Tyrosine Phosphorylation ◽  
Erythrocyte Membrane ◽  
Blood Cells ◽  
Band 3 ◽  
Membrane Modification ◽  
Syk Kinase ◽  
Key Points ◽  
Parasite Egress

Key PointsInhibitors of human Syk kinase suppress parasite egress. Syk inhibitors prevent the tyrosine phosphorylation of band 3 in P falciparum parasitized red blood cells, reducing the release of microparticles.

Two native types of phytochrome A, phyAʹ and phyAʺ, differ by the state of phosphorylation at the N-terminus as revealed by fluorescence investigations of the Ser/Ala mutant of rice phyA expressed in transgenic Arabidopsis

2018 ◽  
Vol 45 (2) ◽  
pp. 150 ◽  
Author(s):  
Vitaly A. Sineshchekov ◽  
Larissa A. Koppel ◽  
Cordelia Bolle
Keyword(s):  
Transgenic Arabidopsis ◽  
Emission Spectra ◽  
Total Content ◽  
The State ◽  
Serine Phosphorylation ◽  
Phytochrome A ◽  
Wild Type ◽  
Etiolated Seedlings ◽  
Pre Treatment

Phytochrome A (phyA) mediates different photoresponses what may be connected with the existence of its two types, phyAʹ and phyAʹʹ, differing by spectroscopic, photochemical and functional properties. We investigated a role of phyA phosphorylation in their formation turning to transgenic Arabidopsis thaliana (L. Heynh.) phyA or phyAphyB mutants overexpressing rice wild-type phyA (phyA WT) or mutant phyA (phyA SA) with the first 10 serines substituted by alanines. This prevents phyA phosphorylation at these sites and modifies photoresponses. Etiolated seedlings were employed and phyA parameters were evaluated with the use of low temperature fluorescence spectroscopy and photochemistry. Germination of seeds was induced by white light (WL) pre-treatment for 15 min or 3 h. Emission spectra of rice phyA WT and phyA SA were similar and their total content was comparable. However, the phyAʹ/phyAʹʹ proportion in phyA WT was high and varied with the duration of the WL pre-treatment, whereas in phyA SA it was substantially shifted towards phyAʹʹ and did not depend on the pre-illumination. This suggests that phyA SA comprises primarily or exclusively the phyAʹʹ pool and supports the notion that the two phyA types differ by the state of serine phosphorylation. phyAʹʹ was also found to be much more effective in the germination induction than phyAʹ.

Sign in / Sign up

Export Citation Format

Share Document