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

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. © 2019, Vögtle et al.

Original language	English
Article number	e46840
Number of pages	43
Journal	eLife
Volume	8
DOIs	https://doi.org/10.7554/eLife.46840
Publication status	Published - 22 Aug 2019

Keywords

parasites and microbes
staffpaper

Access to Document

10.7554/eLife.46840

elife-46840-v2Final published version, 4.01 MBLicence: CC BY

Cite this

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{8d48abcea8ee4c08b8722d5b1770f591,

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. {\textcopyright} 2019, V{\"o}gtle et al.",

keywords = "parasites and microbes, staffpaper",

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}",

year = "2019",

month = aug,

day = "22",

doi = "10.7554/eLife.46840",

language = "English",

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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

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. © 2019, Vögtle et al.

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. © 2019, Vögtle et al.

KW - parasites and microbes

KW - staffpaper

U2 - 10.7554/eLife.46840

DO - 10.7554/eLife.46840

M3 - Article

SN - 2050-084X

VL - 8

JO - eLife

JF - eLife

M1 - e46840

ER -