Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B

Timo Vögtle, Sumana Sharma, Jun Mori, Zoltan Nagy, Daniela Semeniak, Cyril Scandola, Mitchell J. Geer, Christopher W. Smith, Jordan Lane, Scott Pollack, Riitta Lassila, Annukka Jouppila, Alastair J. Barr, Derek J. Ogg, Tina D. Howard, Helen J. McMiken, Juli Warwicker, Catherine Geh, Rachel Rowlinson, W. Mark AbbottAnita Eckly, Harald Schulze, Gavin J. Wright, Alexandra Mazharian, Klaus Fütterer, Sundaresan Rajesh, Michael R. Douglas, Yotis A. Senis

Research output: Contribution to journalArticle

Abstract

The immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor G6b-B is critical for platelet production and activation. Loss of G6b-B results in severe macrothrombocytopenia, myelofibrosis and aberrant platelet function in mice and humans. Using a combination of immunohistochemistry, affinity chromatography and proteomics, we identified the extracellular matrix heparan sulfate (HS) proteoglycan perlecan as a G6b-B binding partner. Subsequent in vitro biochemical studies and a cell-based genetic screen demonstrated that the interaction is specifically mediated by the HS chains of perlecan. Biophysical analysis revealed that heparin forms a high-affinity complex with G6b-B and mediates dimerization. Using platelets from humans and genetically modified mice, we demonstrate that binding of G6b-B to HS and multivalent heparin inhibits platelet and megakaryocyte function by inducing downstream signaling via the tyrosine phosphatases Shp1 and Shp2. Our findings provide novel insights into how G6b-B is regulated and contribute to our understanding of the interaction of megakaryocytes and platelets with glycans.

Original languageEnglish
Article numbere46840
JournaleLife
Volume8
DOIs
Publication statusPublished - 22 Aug 2019

Fingerprint

Megakaryocytes
Heparitin Sulfate
Platelets
Blood Platelets
Immunoreceptor Tyrosine-Based Inhibition Motif
Heparin
Tyrosine
Heparan Sulfate Proteoglycans
Primary Myelofibrosis
Platelet Activation
Dimerization
Affinity chromatography
Affinity Chromatography
Phosphoric Monoester Hydrolases
Proteomics
Extracellular Matrix
Polysaccharides
Immunohistochemistry
Chemical activation
perlecan

Bibliographical note

Copyright Vogtle et al. This
article is distributed under the
terms of the Creative Commons
Attribution License, which
permits unrestricted use and
redistribution provided that the
original author and source are
credited.

Keywords

  • biochemistry
  • cell biology
  • chemical biology
  • G6b-B
  • heparan sulfate
  • heparin
  • human
  • ITIM-receptor
  • mouse
  • perlecan
  • platelets

Cite this

Vögtle, T., Sharma, S., Mori, J., Nagy, Z., Semeniak, D., Scandola, C., ... Senis, Y. A. (2019). Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B. eLife, 8, [e46840]. https://doi.org/10.7554/eLife.46840
Vögtle, Timo ; Sharma, Sumana ; Mori, Jun ; Nagy, Zoltan ; Semeniak, Daniela ; Scandola, Cyril ; Geer, Mitchell J. ; Smith, Christopher W. ; Lane, Jordan ; Pollack, Scott ; Lassila, Riitta ; Jouppila, Annukka ; Barr, Alastair J. ; Ogg, Derek J. ; Howard, Tina D. ; McMiken, Helen J. ; Warwicker, Juli ; Geh, Catherine ; Rowlinson, Rachel ; Abbott, W. Mark ; Eckly, Anita ; Schulze, Harald ; Wright, Gavin J. ; Mazharian, Alexandra ; Fütterer, Klaus ; Rajesh, Sundaresan ; Douglas, Michael R. ; Senis, Yotis A. / Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B. In: eLife. 2019 ; Vol. 8.
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abstract = "The immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor G6b-B is critical for platelet production and activation. Loss of G6b-B results in severe macrothrombocytopenia, myelofibrosis and aberrant platelet function in mice and humans. Using a combination of immunohistochemistry, affinity chromatography and proteomics, we identified the extracellular matrix heparan sulfate (HS) proteoglycan perlecan as a G6b-B binding partner. Subsequent in vitro biochemical studies and a cell-based genetic screen demonstrated that the interaction is specifically mediated by the HS chains of perlecan. Biophysical analysis revealed that heparin forms a high-affinity complex with G6b-B and mediates dimerization. Using platelets from humans and genetically modified mice, we demonstrate that binding of G6b-B to HS and multivalent heparin inhibits platelet and megakaryocyte function by inducing downstream signaling via the tyrosine phosphatases Shp1 and Shp2. Our findings provide novel insights into how G6b-B is regulated and contribute to our understanding of the interaction of megakaryocytes and platelets with glycans.",
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Vögtle, T, Sharma, S, Mori, J, Nagy, Z, Semeniak, D, Scandola, C, Geer, MJ, Smith, CW, Lane, J, Pollack, S, Lassila, R, Jouppila, A, Barr, AJ, Ogg, DJ, Howard, TD, McMiken, HJ, Warwicker, J, Geh, C, Rowlinson, R, Abbott, WM, Eckly, A, Schulze, H, Wright, GJ, Mazharian, A, Fütterer, K, Rajesh, S, Douglas, MR & Senis, YA 2019, 'Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B', eLife, vol. 8, e46840. https://doi.org/10.7554/eLife.46840

Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B. / Vögtle, Timo; Sharma, Sumana; Mori, Jun; Nagy, Zoltan; Semeniak, Daniela; Scandola, Cyril; Geer, Mitchell J.; Smith, Christopher W.; Lane, Jordan; Pollack, Scott; Lassila, Riitta; Jouppila, Annukka; Barr, Alastair J.; Ogg, Derek J.; Howard, Tina D.; McMiken, Helen J.; Warwicker, Juli; Geh, Catherine; Rowlinson, Rachel; Abbott, W. Mark; Eckly, Anita; Schulze, Harald; Wright, Gavin J.; Mazharian, Alexandra; Fütterer, Klaus; Rajesh, Sundaresan; Douglas, Michael R.; Senis, Yotis A.

In: eLife, Vol. 8, e46840, 22.08.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B

AU - Vögtle, Timo

AU - Sharma, Sumana

AU - Mori, Jun

AU - Nagy, Zoltan

AU - Semeniak, Daniela

AU - Scandola, Cyril

AU - Geer, Mitchell J.

AU - Smith, Christopher W.

AU - Lane, Jordan

AU - Pollack, Scott

AU - Lassila, Riitta

AU - Jouppila, Annukka

AU - Barr, Alastair J.

AU - Ogg, Derek J.

AU - Howard, Tina D.

AU - McMiken, Helen J.

AU - Warwicker, Juli

AU - Geh, Catherine

AU - Rowlinson, Rachel

AU - Abbott, W. Mark

AU - Eckly, Anita

AU - Schulze, Harald

AU - Wright, Gavin J.

AU - Mazharian, Alexandra

AU - Fütterer, Klaus

AU - Rajesh, Sundaresan

AU - Douglas, Michael R.

AU - Senis, Yotis A.

N1 - Copyright Vogtle et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

PY - 2019/8/22

Y1 - 2019/8/22

N2 - The immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor G6b-B is critical for platelet production and activation. Loss of G6b-B results in severe macrothrombocytopenia, myelofibrosis and aberrant platelet function in mice and humans. Using a combination of immunohistochemistry, affinity chromatography and proteomics, we identified the extracellular matrix heparan sulfate (HS) proteoglycan perlecan as a G6b-B binding partner. Subsequent in vitro biochemical studies and a cell-based genetic screen demonstrated that the interaction is specifically mediated by the HS chains of perlecan. Biophysical analysis revealed that heparin forms a high-affinity complex with G6b-B and mediates dimerization. Using platelets from humans and genetically modified mice, we demonstrate that binding of G6b-B to HS and multivalent heparin inhibits platelet and megakaryocyte function by inducing downstream signaling via the tyrosine phosphatases Shp1 and Shp2. Our findings provide novel insights into how G6b-B is regulated and contribute to our understanding of the interaction of megakaryocytes and platelets with glycans.

AB - The immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptor G6b-B is critical for platelet production and activation. Loss of G6b-B results in severe macrothrombocytopenia, myelofibrosis and aberrant platelet function in mice and humans. Using a combination of immunohistochemistry, affinity chromatography and proteomics, we identified the extracellular matrix heparan sulfate (HS) proteoglycan perlecan as a G6b-B binding partner. Subsequent in vitro biochemical studies and a cell-based genetic screen demonstrated that the interaction is specifically mediated by the HS chains of perlecan. Biophysical analysis revealed that heparin forms a high-affinity complex with G6b-B and mediates dimerization. Using platelets from humans and genetically modified mice, we demonstrate that binding of G6b-B to HS and multivalent heparin inhibits platelet and megakaryocyte function by inducing downstream signaling via the tyrosine phosphatases Shp1 and Shp2. Our findings provide novel insights into how G6b-B is regulated and contribute to our understanding of the interaction of megakaryocytes and platelets with glycans.

KW - biochemistry

KW - cell biology

KW - chemical biology

KW - G6b-B

KW - heparan sulfate

KW - heparin

KW - human

KW - ITIM-receptor

KW - mouse

KW - perlecan

KW - platelets

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DO - 10.7554/eLife.46840

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VL - 8

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Vögtle T, Sharma S, Mori J, Nagy Z, Semeniak D, Scandola C et al. Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B. eLife. 2019 Aug 22;8. e46840. https://doi.org/10.7554/eLife.46840