Platelet protein disulfide isomerase is localized in the dense tubular system and does not become surface expressed after activation

Pannerden, Hezder E. van Nispen tot; Dijk, Suzanne M. van; Du, Vivian X.; Heijnen, Harry F. G.

doi:10.1182/blood-2009-03-210450

Cited by 26 publications

(27 citation statements)

References 15 publications

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“…The soluble cytosolic marker RabGDI was mainly concentrated at the top of the gradient as were the fractions containing calreticulin, a marker for the DTS previously used to detect PDI in this compartment in platelets. 19 TSP-1 was abundantly expressed in the heaviest fractions where α –granules are fractionated. Consistent with our imaging and previous results, 38 PDI and ERp57 were concentrated in the lightest fractions, indicating association with cell-surface and/or near-surface membrane structures such as the DTS.…”

Section: Resultsmentioning

confidence: 95%

“…6, 8, 10, 15-19 In this study we tracked PDI and ERp57 through development of precursor megakaryocytes (MK) and platelets, where we observed that these enzymes are concentrated in a subcellular compartment distinct from known secretory vesicles that overlaps with the dense tubular system (DTS) derived from MK smooth ER, and with the inner surface of the platelet outer membrane where the sarco/endoplasmic reticulum calcium ATPase SERCA3 is known to be localized. 20 During platelet activation, actin polymerization facilitates the movement of secretory vesicles to the cell plasma membrane surface, 21 where the interaction of soluble N-ethylmaleimide-sensitive fusion protein attachment receptors (SNAREs) facilitates membrane fusion mediated by Munc 13-4.…”

Section: Introductionmentioning

confidence: 99%

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Intracellular Trafficking, Localization, and Mobilization of Platelet-Borne Thiol Isomerases

Crescente

Pluthero

et al. 2016

ATVB

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Objective Thiol isomerases facilitate protein folding in the endoplasmic reticulum, and several of these enzymes, including PDI and ERp57, are mobilized to the surface of activated platelets, where they influence platelet aggregation, blood coagulation and thrombus formation. In this study we examined for the first time the synthesis and trafficking of thiol isomerases in megakaryocytes, determined their subcellular localization in platelets and identified the cellular events responsible for their movement to the platelet surface upon activation. Approach and Results Immunofluorescence microscopy (IFM) imaging was used to localize PDI and ERp57 in murine and human megakaryocytes at various developmental stages. IFM and subcellular fractionation analysis were used to localize these proteins in platelets to a compartment distinct from known secretory vesicles that overlaps with an inner cell surface membrane region defined by the endoplasmic/sarcoplasmic reticulum proteins calnexin and SERCA3. IFM and flow cytometry were used to monitor thiol isomerase mobilization in activated platelets in the presence and absence of actin polymerization (inhibited by latrunculin), and in the presence or absence of membrane fusion mediated by Munc 13-4 (absent in platelets from Unc13dJinx mice). Conclusions Platelet-borne thiol isomerases are trafficked independently of secretory granule contents in megakaryocytes, and become concentrated in a subcellular compartment near the inner surface of the platelet outer membrane corresponding to the sarco/endoplasmic reticulum of these cells. Thiol isomerases are mobilized to the surface of activated platelets via a process that requires actin polymerization but not SNARE/Munc 13-4-dependent vesicular-plasma membrane fusion.

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Section: Resultsmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Intracellular Trafficking, Localization, and Mobilization of Platelet-Borne Thiol Isomerases

Crescente

Pluthero

et al. 2016

ATVB

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“…Both microscopy 48, 49 and density gradient centrifugation 9 show that endothelial PDI is concentrated in the ER. Several experiments, however, indicate that PDI also resides on the surface of endothelial cells.…”

Section: Introductionmentioning

confidence: 99%

Thiol Isomerases in Thrombus Formation

Furie

Flaumenhaft

2014

Circulation Research

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Protein disulfide isomerase, ERp5 and ERp57, among perhaps other thiol isomerases, are important for the initiation of thrombus formation. Using the laser injury thrombosis model in mice to induce in vivo arterial thrombus formation, it was shown that thrombus formation is associated with PDI secretion by platelets, that inhibition of PDI blocked platelet thrombus formation and fibrin generation, and that endothelial cell activation leads to PDI secretion. Similar results using this and other thrombosis models in mice have demonstrated the importance of ERp5 and ERp57 in the initiation of thrombus formation. The integrins αIIbβ3 and αVβ3 play a key role in this process and interact directly with PDI, ERp5 and ERp57. The mechanism by which thiol isomerases participate in thrombus generation is being evaluated using trapping mutant forms to identify substrates of thiol isomerases that participate in the network pathways linking thiol isomerases, platelet receptor activation and fibrin generation. Protein disulfide isomerase as an antithrombotic target is being explored using isoquercetin and quercetin 3-rutinoside, inhibitors of PDI identified by high throughput screening. Regulation of thiol isomerase expression, analysis of the storage and secretion of thiol isomerases and determination of the electron transfer pathway are key issues to understanding this newly discovered mechanism of regulation of the initiation of thrombus formation.

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“…3 These rapid membrane dynamics are crucial for the progression of platelet-substrate interaction (spreading) and subsequent plateletplatelet interaction (aggregation), ultimately leading to the formation of a platelet plug and the arrest of bleeding. 4 Platelets contain several distinct membrane systems: (1) the open canalicular system (OCS), which is continuous with the cell surface and serves as a membrane reservoir during shape change and spreading 5,6 ; (2) the dense tubular system (DTS), representing the platelet smooth endoplasmic reticulum 7 ; and (3) secretory organelles. Four types of secretory organelles have been identified in platelets, based on their ultrastructure and selective protein composition: ␣-granules, dense granules, multivesicular bodies, and lysosomes.…”

Section: Introductionmentioning

confidence: 99%

The platelet interior revisited: electron tomography reveals tubular α-granule subtypes

Pannerden

Haas²,

Geerts

et al. 2010

Blood

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159

136

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We have used (cryo) electron tomography to provide a 3-dimensional (3D) map of the intracellular membrane organization of human platelets at high spatial resolution. Our study shows that the open canalicular system and dense tubular system are highly intertwined and form close associations in specialized membrane regions. 3D reconstructions of individual ␣-granules revealed large heterogeneity in their membrane organization. On the basis of their divergent morphology, we categorized ␣-granules into the following subtypes: spherical granules with electron-dense and electron-lucent zone containing 12-nm von Willebrand factor tubules, subtypes containing a multitude of luminal vesicles, 50-nm-wide tubular organelles, and a population with 18.4-nm crystalline cross-striations. Low-dose (cryo) electron tomography and 3D reconstruction of whole vitrified platelets confirmed the existence of long tubular granules with a remarkably curved architecture. Immunoelectron microscopy confirmed that these extended structures represent ␣-granule subtypes. Tubular ␣-granules represent approximately 16% of the total ␣-granule population and are detected in approximately half of the platelet population. They express membrane-bound proteins GLUT3 and ␣IIb-␤3 integrin and contain abundant fibrinogen and albumin but low levels of ␤-thromboglobulin and no von Willebrand factor. Our 3D study demonstrates that, besides the existence of morphologically different ␣-granule subtypes, high spatial segregation of cargo exists within individual ␣-granules. (Blood. 2010;116(7):1147-1156) IntroductionBlood platelets are the smallest cells in our circulation. They play a central role in the arrest of bleeding after damage of a blood vessel and are crucial elements in the development of thrombosis. 1,2 On injury, platelets rapidly adhere to components of the subendothelium, followed by shape change and subsequent granule secretion. 3 These rapid membrane dynamics are crucial for the progression of platelet-substrate interaction (spreading) and subsequent plateletplatelet interaction (aggregation), ultimately leading to the formation of a platelet plug and the arrest of bleeding. 4 Platelets contain several distinct membrane systems: (1) the open canalicular system (OCS), which is continuous with the cell surface and serves as a membrane reservoir during shape change and spreading 5,6 ; (2) the dense tubular system (DTS), representing the platelet smooth endoplasmic reticulum 7 ; and (3) secretory organelles. Four types of secretory organelles have been identified in platelets, based on their ultrastructure and selective protein composition: ␣-granules, dense granules, multivesicular bodies, and lysosomes. 8-10 ␣-Granules are the major secretory organelles and appear in electron microscopy cross sections as 200-to 500-nm spherical organelles. Platelet ␣-granules and dense granules are differentially released and play crucial roles in the secondary platelet response. 11,12 Recent studies have suggested the existence of ␣-granule subclasses with dif...

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Platelet protein disulfide isomerase is localized in the dense tubular system and does not become surface expressed after activation

Cited by 26 publications

References 15 publications

Intracellular Trafficking, Localization, and Mobilization of Platelet-Borne Thiol Isomerases

Intracellular Trafficking, Localization, and Mobilization of Platelet-Borne Thiol Isomerases

Thiol Isomerases in Thrombus Formation

The platelet interior revisited: electron tomography reveals tubular α-granule subtypes

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