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 Abbott; Anita Eckly; Harald Schulze; Gavin J. Wright; Alexandra Mazharian; Klaus Fütterer; Sundaresan Rajesh; Michael R. Douglas; Yotis A. Senis

doi:10.7554/eLife.46840

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

Aston Pharmacy School

Research output: Contribution to journal › Article › peer-review

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 language	English
Article number	e46840
Journal	eLife
Volume	8
DOIs	https://doi.org/10.7554/eLife.46840
Publication status	Published - 22 Aug 2019

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

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10.7554/eLife.46840Licence: CC BY 3.0

Heparan sulfates are critical regulatorsFinal published version, 4.01 MBLicence: CC BY 3.0

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Vögtle, T., Sharma, S., Mori, J., Nagy, Z., Semeniak, D., Scandola, C., Geer, M. J., Smith, C. W., Lane, J., Pollack, S., Lassila, R., Jouppila, A., Barr, A. J., Ogg, D. J., Howard, T. D., McMiken, H. J., Warwicker, J., Geh, C., Rowlinson, R., ... Senis, Y. A. (2019). Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B. eLife, 8, Article e46840. https://doi.org/10.7554/eLife.46840

@article{635434836d7d4f0093a14bf6299140f8,

title = "Heparan sulfates are critical regulators of the inhibitory megakaryocyte-platelet receptor G6b-B",

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.",

keywords = "biochemistry, cell biology, chemical biology, G6b-B, heparan sulfate, heparin, human, ITIM-receptor, mouse, perlecan, platelets",

author = "Timo V{\"o}gtle and Sumana Sharma and Jun Mori and Zoltan Nagy and Daniela Semeniak and Cyril Scandola and Geer, {Mitchell J.} and Smith, {Christopher W.} and Jordan Lane and Scott Pollack and Riitta Lassila and Annukka Jouppila and Barr, {Alastair J.} and Ogg, {Derek J.} and Howard, {Tina D.} and McMiken, {Helen J.} and Juli Warwicker and Catherine Geh and Rachel Rowlinson and Abbott, {W. Mark} and Anita Eckly and Harald Schulze and Wright, {Gavin J.} and Alexandra Mazharian and Klaus F{\"u}tterer and Sundaresan Rajesh and Douglas, {Michael R.} and Senis, {Yotis A.}",

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. ",

year = "2019",

month = aug,

day = "22",

doi = "10.7554/eLife.46840",

language = "English",

volume = "8",

journal = "eLife",

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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

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

UR - http://www.scopus.com/inward/record.url?scp=85072509074&partnerID=8YFLogxK

UR - https://elifesciences.org/articles/46840

U2 - 10.7554/eLife.46840

DO - 10.7554/eLife.46840

M3 - Article

C2 - 31436532

AN - SCOPUS:85072509074

SN - 2050-084X

VL - 8

JO - eLife

JF - eLife

M1 - e46840

ER -

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

Abstract

Bibliographical note

Keywords

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