A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation

Hiromi Watari; Hiromu Kageyama; Nami Masubuchi; Hiroya Nakajima; Kako Onodera; Pamela J. Focia; Takumi Oshiro; Takashi Matsui; Yoshio Kodera; Tomohisa Ogawa; Takeshi Yokoyama; Makoto Hirayama; Kanji Hori; Douglas M. Freymann; Misa Imai; Norio Komatsu; Marito Araki; Yoshikazu Tanaka; Ryuichi Sakai

doi:10.1038/s41467-022-34921-2

A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation

Hiromi Watari, Hiromu Kageyama, Nami Masubuchi, Hiroya Nakajima, Kako Onodera, Pamela J. Focia, Takumi Oshiro, Takashi Matsui, Yoshio Kodera, Tomohisa Ogawa, Takeshi Yokoyama, Makoto Hirayama, Kanji Hori, Douglas M. Freymann, Misa Imai, Norio Komatsu, Marito Araki^*, Yoshikazu Tanaka^*, Ryuichi Sakai^*

^*Corresponding author for this work

Biochemistry and Molecular Genetics

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

1 Scopus citations

Abstract

N-glycan-mediated activation of the thrombopoietin receptor (MPL) under pathological conditions has been implicated in myeloproliferative neoplasms induced by mutant calreticulin, which forms an endogenous receptor-agonist complex that traffics to the cell surface and constitutively activates the receptor. However, the molecular basis for this mechanism is elusive because oncogenic activation occurs only in the cell-intrinsic complex and is thus cannot be replicated with external agonists. Here, we describe the structure and function of a marine sponge-derived MPL agonist, thrombocorticin (ThC), a homodimerized lectin with calcium-dependent fucose-binding properties. In-depth characterization of lectin-induced activation showed that, similar to oncogenic activation, sugar chain-mediated activation persists due to limited receptor internalization. The strong synergy between ThC and thrombopoietin suggests that ThC catalyzes the formation of receptor dimers on the cell surface. Overall, the existence of sugar-mediated MPL activation, in which the mode of activation is different from the original ligand, suggests that receptor activation is unpredictably diverse in living organisms.

Original language	English (US)
Article number	7262
Journal	Nature communications
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41467-022-34921-2
State	Published - Dec 2022

Funding

We thank Professor Yasuhiko Kizuka at Gifu University for providing 6-alkynyl fucose and valuable comments. We also thank Dr. Yinjie Yang and Ms. Mai Nudejima for their technical assistances, Dr. Takanori Nakamura at Nissan Chemical Co. Ltd. and Dr. Megumi Funakoshi-Tago at Faculty of Pharmacy, Keio University for providing Ba/F3 cells, the Global Facility Center at Hokkaido University for performing amino acid sequence analysis, the Laboratory of Molecular and Biochemical Research and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine for their technical assistants for immunoblot and FACS analysis, the Teijin Scholarship Foundation, Suntory Foundation for Life Science and the Japan Society for the Promotion of Science (JSPS) DC2 for scholarships to H.W. Japan Society for the Promotion of Science (JSPS) #19H03040 (R.S.), #22H02430 (R.S.), #21K08376 (M.I.), #21K08405 (N.M.), #21K08424 (N.K.), #19K08848 (M.A.), #22H02252 (Y.T.), #22H02915 (Y.T.), #22K20589 (H.W.), and Grant-in-aid for JSPS fellows #20J11377(H.W.) for fundings. We also thank funding from Ikeda Scientific Co., Ltd (H.W.), the SENSHIN Medical Research Foundation (M.A.) and Takeda Science Foundation (M.A.). We are grateful to Chuuk State Department of Marine Resources for providing permission to collect sponges. We thank Professor Yasuhiko Kizuka at Gifu University for providing 6-alkynyl fucose and valuable comments. We also thank Dr. Yinjie Yang and Ms. Mai Nudejima for their technical assistances, Dr. Takanori Nakamura at Nissan Chemical Co. Ltd. and Dr. Megumi Funakoshi-Tago at Faculty of Pharmacy, Keio University for providing Ba/F3 cells, the Global Facility Center at Hokkaido University for performing amino acid sequence analysis, the Laboratory of Molecular and Biochemical Research and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine for their technical assistants for immunoblot and FACS analysis, the Teijin Scholarship Foundation, Suntory Foundation for Life Science and the Japan Society for the Promotion of Science (JSPS) DC2 for scholarships to H.W. Japan Society for the Promotion of Science (JSPS) #19H03040 (R.S.), #22H02430 (R.S.), #21K08376 (M.I.), #21K08405 (N.M.), #21K08424 (N.K.), #19K08848 (M.A.), #22H02252 (Y.T.), #22H02915 (Y.T.), #22K20589 (H.W.), and Grant-in-aid for JSPS fellows #20J11377(H.W.) for fundings. We also thank funding from Ikeda Scientific Co., Ltd (H.W.), the SENSHIN Medical Research Foundation (M.A.) and Takeda Science Foundation (M.A.). We are grateful to Chuuk State Department of Marine Resources for providing permission to collect sponges.

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1038/s41467-022-34921-2

Cite this

Watari, H., Kageyama, H., Masubuchi, N., Nakajima, H., Onodera, K., Focia, P. J., Oshiro, T., Matsui, T., Kodera, Y., Ogawa, T., Yokoyama, T., Hirayama, M., Hori, K., Freymann, D. M., Imai, M., Komatsu, N., Araki, M., Tanaka, Y., & Sakai, R. (2022). A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation. Nature communications, 13(1), Article 7262. https://doi.org/10.1038/s41467-022-34921-2

@article{b61efd5d9bc54e1ea5139f32e55a4b08,

title = "A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation",

abstract = "N-glycan-mediated activation of the thrombopoietin receptor (MPL) under pathological conditions has been implicated in myeloproliferative neoplasms induced by mutant calreticulin, which forms an endogenous receptor-agonist complex that traffics to the cell surface and constitutively activates the receptor. However, the molecular basis for this mechanism is elusive because oncogenic activation occurs only in the cell-intrinsic complex and is thus cannot be replicated with external agonists. Here, we describe the structure and function of a marine sponge-derived MPL agonist, thrombocorticin (ThC), a homodimerized lectin with calcium-dependent fucose-binding properties. In-depth characterization of lectin-induced activation showed that, similar to oncogenic activation, sugar chain-mediated activation persists due to limited receptor internalization. The strong synergy between ThC and thrombopoietin suggests that ThC catalyzes the formation of receptor dimers on the cell surface. Overall, the existence of sugar-mediated MPL activation, in which the mode of activation is different from the original ligand, suggests that receptor activation is unpredictably diverse in living organisms.",

author = "Hiromi Watari and Hiromu Kageyama and Nami Masubuchi and Hiroya Nakajima and Kako Onodera and Focia, {Pamela J.} and Takumi Oshiro and Takashi Matsui and Yoshio Kodera and Tomohisa Ogawa and Takeshi Yokoyama and Makoto Hirayama and Kanji Hori and Freymann, {Douglas M.} and Misa Imai and Norio Komatsu and Marito Araki and Yoshikazu Tanaka and Ryuichi Sakai",

note = "Funding Information: We thank Professor Yasuhiko Kizuka at Gifu University for providing 6-alkynyl fucose and valuable comments. We also thank Dr. Yinjie Yang and Ms. Mai Nudejima for their technical assistances, Dr. Takanori Nakamura at Nissan Chemical Co. Ltd. and Dr. Megumi Funakoshi-Tago at Faculty of Pharmacy, Keio University for providing Ba/F3 cells, the Global Facility Center at Hokkaido University for performing amino acid sequence analysis, the Laboratory of Molecular and Biochemical Research and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine for their technical assistants for immunoblot and FACS analysis, the Teijin Scholarship Foundation, Suntory Foundation for Life Science and the Japan Society for the Promotion of Science (JSPS) DC2 for scholarships to H.W. Japan Society for the Promotion of Science (JSPS) #19H03040 (R.S.), #22H02430 (R.S.), #21K08376 (M.I.), #21K08405 (N.M.), #21K08424 (N.K.), #19K08848 (M.A.), #22H02252 (Y.T.), #22H02915 (Y.T.), #22K20589 (H.W.), and Grant-in-aid for JSPS fellows #20J11377(H.W.) for fundings. We also thank funding from Ikeda Scientific Co., Ltd (H.W.), the SENSHIN Medical Research Foundation (M.A.) and Takeda Science Foundation (M.A.). We are grateful to Chuuk State Department of Marine Resources for providing permission to collect sponges. Funding Information: We thank Professor Yasuhiko Kizuka at Gifu University for providing 6-alkynyl fucose and valuable comments. We also thank Dr. Yinjie Yang and Ms. Mai Nudejima for their technical assistances, Dr. Takanori Nakamura at Nissan Chemical Co. Ltd. and Dr. Megumi Funakoshi-Tago at Faculty of Pharmacy, Keio University for providing Ba/F3 cells, the Global Facility Center at Hokkaido University for performing amino acid sequence analysis, the Laboratory of Molecular and Biochemical Research and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine for their technical assistants for immunoblot and FACS analysis, the Teijin Scholarship Foundation, Suntory Foundation for Life Science and the Japan Society for the Promotion of Science (JSPS) DC2 for scholarships to H.W. Japan Society for the Promotion of Science (JSPS) #19H03040 (R.S.), #22H02430 (R.S.), #21K08376 (M.I.), #21K08405 (N.M.), #21K08424 (N.K.), #19K08848 (M.A.), #22H02252 (Y.T.), #22H02915 (Y.T.), #22K20589 (H.W.), and Grant-in-aid for JSPS fellows #20J11377(H.W.) for fundings. We also thank funding from Ikeda Scientific Co., Ltd (H.W.), the SENSHIN Medical Research Foundation (M.A.) and Takeda Science Foundation (M.A.). We are grateful to Chuuk State Department of Marine Resources for providing permission to collect sponges. Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = dec,

doi = "10.1038/s41467-022-34921-2",

language = "English (US)",

volume = "13",

journal = "Nature communications",

issn = "2041-1723",

publisher = "Nature Research",

number = "1",

}

Watari, H, Kageyama, H, Masubuchi, N, Nakajima, H, Onodera, K, Focia, PJ, Oshiro, T, Matsui, T, Kodera, Y, Ogawa, T, Yokoyama, T, Hirayama, M, Hori, K, Freymann, DM, Imai, M, Komatsu, N, Araki, M, Tanaka, Y & Sakai, R 2022, 'A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation', Nature communications, vol. 13, no. 1, 7262. https://doi.org/10.1038/s41467-022-34921-2

TY - JOUR

T1 - A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation

AU - Watari, Hiromi

AU - Kageyama, Hiromu

AU - Masubuchi, Nami

AU - Nakajima, Hiroya

AU - Onodera, Kako

AU - Focia, Pamela J.

AU - Oshiro, Takumi

AU - Matsui, Takashi

AU - Kodera, Yoshio

AU - Ogawa, Tomohisa

AU - Yokoyama, Takeshi

AU - Hirayama, Makoto

AU - Hori, Kanji

AU - Freymann, Douglas M.

AU - Imai, Misa

AU - Komatsu, Norio

AU - Araki, Marito

AU - Tanaka, Yoshikazu

AU - Sakai, Ryuichi

N1 - Funding Information: We thank Professor Yasuhiko Kizuka at Gifu University for providing 6-alkynyl fucose and valuable comments. We also thank Dr. Yinjie Yang and Ms. Mai Nudejima for their technical assistances, Dr. Takanori Nakamura at Nissan Chemical Co. Ltd. and Dr. Megumi Funakoshi-Tago at Faculty of Pharmacy, Keio University for providing Ba/F3 cells, the Global Facility Center at Hokkaido University for performing amino acid sequence analysis, the Laboratory of Molecular and Biochemical Research and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine for their technical assistants for immunoblot and FACS analysis, the Teijin Scholarship Foundation, Suntory Foundation for Life Science and the Japan Society for the Promotion of Science (JSPS) DC2 for scholarships to H.W. Japan Society for the Promotion of Science (JSPS) #19H03040 (R.S.), #22H02430 (R.S.), #21K08376 (M.I.), #21K08405 (N.M.), #21K08424 (N.K.), #19K08848 (M.A.), #22H02252 (Y.T.), #22H02915 (Y.T.), #22K20589 (H.W.), and Grant-in-aid for JSPS fellows #20J11377(H.W.) for fundings. We also thank funding from Ikeda Scientific Co., Ltd (H.W.), the SENSHIN Medical Research Foundation (M.A.) and Takeda Science Foundation (M.A.). We are grateful to Chuuk State Department of Marine Resources for providing permission to collect sponges. Funding Information: We thank Professor Yasuhiko Kizuka at Gifu University for providing 6-alkynyl fucose and valuable comments. We also thank Dr. Yinjie Yang and Ms. Mai Nudejima for their technical assistances, Dr. Takanori Nakamura at Nissan Chemical Co. Ltd. and Dr. Megumi Funakoshi-Tago at Faculty of Pharmacy, Keio University for providing Ba/F3 cells, the Global Facility Center at Hokkaido University for performing amino acid sequence analysis, the Laboratory of Molecular and Biochemical Research and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine for their technical assistants for immunoblot and FACS analysis, the Teijin Scholarship Foundation, Suntory Foundation for Life Science and the Japan Society for the Promotion of Science (JSPS) DC2 for scholarships to H.W. Japan Society for the Promotion of Science (JSPS) #19H03040 (R.S.), #22H02430 (R.S.), #21K08376 (M.I.), #21K08405 (N.M.), #21K08424 (N.K.), #19K08848 (M.A.), #22H02252 (Y.T.), #22H02915 (Y.T.), #22K20589 (H.W.), and Grant-in-aid for JSPS fellows #20J11377(H.W.) for fundings. We also thank funding from Ikeda Scientific Co., Ltd (H.W.), the SENSHIN Medical Research Foundation (M.A.) and Takeda Science Foundation (M.A.). We are grateful to Chuuk State Department of Marine Resources for providing permission to collect sponges. Publisher Copyright: © 2022, The Author(s).

PY - 2022/12

Y1 - 2022/12

N2 - N-glycan-mediated activation of the thrombopoietin receptor (MPL) under pathological conditions has been implicated in myeloproliferative neoplasms induced by mutant calreticulin, which forms an endogenous receptor-agonist complex that traffics to the cell surface and constitutively activates the receptor. However, the molecular basis for this mechanism is elusive because oncogenic activation occurs only in the cell-intrinsic complex and is thus cannot be replicated with external agonists. Here, we describe the structure and function of a marine sponge-derived MPL agonist, thrombocorticin (ThC), a homodimerized lectin with calcium-dependent fucose-binding properties. In-depth characterization of lectin-induced activation showed that, similar to oncogenic activation, sugar chain-mediated activation persists due to limited receptor internalization. The strong synergy between ThC and thrombopoietin suggests that ThC catalyzes the formation of receptor dimers on the cell surface. Overall, the existence of sugar-mediated MPL activation, in which the mode of activation is different from the original ligand, suggests that receptor activation is unpredictably diverse in living organisms.

AB - N-glycan-mediated activation of the thrombopoietin receptor (MPL) under pathological conditions has been implicated in myeloproliferative neoplasms induced by mutant calreticulin, which forms an endogenous receptor-agonist complex that traffics to the cell surface and constitutively activates the receptor. However, the molecular basis for this mechanism is elusive because oncogenic activation occurs only in the cell-intrinsic complex and is thus cannot be replicated with external agonists. Here, we describe the structure and function of a marine sponge-derived MPL agonist, thrombocorticin (ThC), a homodimerized lectin with calcium-dependent fucose-binding properties. In-depth characterization of lectin-induced activation showed that, similar to oncogenic activation, sugar chain-mediated activation persists due to limited receptor internalization. The strong synergy between ThC and thrombopoietin suggests that ThC catalyzes the formation of receptor dimers on the cell surface. Overall, the existence of sugar-mediated MPL activation, in which the mode of activation is different from the original ligand, suggests that receptor activation is unpredictably diverse in living organisms.

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U2 - 10.1038/s41467-022-34921-2

DO - 10.1038/s41467-022-34921-2

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C2 - 36433967

AN - SCOPUS:85142476940

SN - 2041-1723

VL - 13

JO - Nature communications

JF - Nature communications

IS - 1

M1 - 7262

ER -

A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation

Abstract

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ASJC Scopus subject areas

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Thrombocorticin Q25K in complex with Ca2+

Thrombocorticin in complex with Ca2+ and mannose

Thrombocorticin

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A marine sponge-derived lectin reveals hidden pathway for thrombopoietin receptor activation

Abstract

Funding

ASJC Scopus subject areas

UN SDGs

Access to Document

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Fingerprint

Datasets

Thrombocorticin Q25K in complex with Ca2+

Thrombocorticin in complex with Ca2+ and mannose

Thrombocorticin

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