Brain glycogen serves as a critical glucosamine cache required for protein glycosylation

Ramon C. Sun; Lyndsay E.A. Young; Ronald C. Bruntz; Kia H. Markussen; Zhengqiu Zhou; Lindsey R. Conroy; Tara R. Hawkinson; Harrison A. Clarke; Alexandra E. Stanback; Jessica K.A. Macedo; Shane Emanuelle; M. Kathryn Brewer; Alberto L. Rondon; Annette Mestas; William C. Sanders; Krishna K. Mahalingan; Buyun Tang; Vimbai M. Chikwana; Dyann M. Segvich; Christopher J. Contreras; Elizabeth J. Allenger; Christine F. Brainson; Lance A. Johnson; Richard E. Taylor; Dustin D. Armstrong; Robert Shaffer; Charles J. Waechter; Craig W. Vander Kooi; Anna A. DePaoli-Roach; Peter J. Roach; Thomas D. Hurley; Richard R. Drake; Matthew S. Gentry

doi:10.1016/j.cmet.2021.05.003

Brain glycogen serves as a critical glucosamine cache required for protein glycosylation

Ramon C. Sun, Lyndsay E.A. Young, Ronald C. Bruntz, Kia H. Markussen, Zhengqiu Zhou, Lindsey R. Conroy, Tara R. Hawkinson, Harrison A. Clarke, Alexandra E. Stanback, Jessica K.A. Macedo, Shane Emanuelle, M. Kathryn Brewer, Alberto L. Rondon, Annette Mestas, William C. Sanders, Krishna K. Mahalingan, Buyun Tang, Vimbai M. Chikwana, Dyann M. Segvich, Christopher J. ContrerasElizabeth J. Allenger, Christine F. Brainson, Lance A. Johnson, Richard E. Taylor, Dustin D. Armstrong, Robert Shaffer, Charles J. Waechter, Craig W. Vander Kooi, Anna A. DePaoli-Roach, Peter J. Roach, Thomas D. Hurley, Richard R. Drake, Matthew S. Gentry

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

18 Scopus citations

Abstract

Glycosylation defects are a hallmark of many nervous system diseases. However, the molecular and metabolic basis for this pathology is not fully understood. In this study, we found that N-linked protein glycosylation in the brain is metabolically channeled to glucosamine metabolism through glycogenolysis. We discovered that glucosamine is an abundant constituent of brain glycogen, which functions as a glucosamine reservoir for multiple glycoconjugates. We demonstrated the enzymatic incorporation of glucosamine into glycogen by glycogen synthase, and the release by glycogen phosphorylase by biochemical and structural methodologies, in primary astrocytes, and in vivo by isotopic tracing and mass spectrometry. Using two mouse models of glycogen storage diseases, we showed that disruption of brain glycogen metabolism causes global decreases in free pools of UDP-N-acetylglucosamine and N-linked protein glycosylation. These findings revealed fundamental biological roles of brain glycogen in protein glycosylation with direct relevance to multiple human diseases of the central nervous system.

Original language	English
Pages (from-to)	1404-1417.e9
Journal	Cell Metabolism
Volume	33
Issue number	7
DOIs	https://doi.org/10.1016/j.cmet.2021.05.003
State	Published - Jul 6 2021

Bibliographical note

Funding Information:
We would like to thank Dr. Yokota for providing the anti-PGB antibody, Vander Kooi and Gentry lab members for vigorous discussions regarding the work, Mrs. Dana Napier for performing immunohistochemistry on tissue slices, and the Markey Cancer Center. The authors thank Nancy R. Gough (BioSerendipity, LLC) for editorial support. This study was supported by National Institute of Health (NIH) grants R35 NS116824 and P01 NS097197 to M.S.G.; NIH grant R01 AG066653 , St Baldrick’s Career Development Award, V-Scholar Grant, and Rally Foundation Independent Investigator grant to R.C.S.; R01 DK27221 to P.J.R.; and NIH 5R01AG06255002 to L.A.J. J.K.A.M. and L.R.C. were supported by NIH / NCI training grant T32CA165990 . This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH -funded biospecimen Procurement & Translational Pathology Shared Resource Facility of the University of Kentucky Markey Cancer Center ( P30CA177558 ). We also acknowledge partial support of this work through the University of Notre Dame Reisenauer Family GSD Research Fund.

Funding Information:
We would like to thank Dr. Yokota for providing the anti-PGB antibody, Vander Kooi and Gentry lab members for vigorous discussions regarding the work, Mrs. Dana Napier for performing immunohistochemistry on tissue slices, and the Markey Cancer Center. The authors thank Nancy R. Gough (BioSerendipity, LLC) for editorial support. This study was supported by National Institute of Health (NIH) grants R35 NS116824 and P01 NS097197 to M.S.G.; NIH grant R01 AG066653, St Baldrick's Career Development Award, V-Scholar Grant, and Rally Foundation Independent Investigator grant to R.C.S.; R01 DK27221 to P.J.R.; and NIH 5R01AG06255002 to L.A.J. J.K.A.M. and L.R.C. were supported by NIH/NCI training grant T32CA165990. This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH-funded biospecimen Procurement & Translational Pathology Shared Resource Facility of the University of Kentucky Markey Cancer Center (P30CA177558). We also acknowledge partial support of this work through the University of Notre Dame Reisenauer Family GSD Research Fund. Conceptualization, R.C.S. and M.S.G.; methodology, R.C.S. M.S.G. and R.R.D.; investigation, R.C.S. L.E.A.Y. A.E.S. R.C.B. K.H.M. Z.Q.Z. J.K.A.M. S.E. M.K.B. A.L.R. W.C.S. K.K.M. B.T. V.M.C. D.M.S. C.J.C. E.J.A. C.F.B. L.A.J. R.E.T. C.W.V.K. C.W. A.A.D.R. P.J.R. T.D.H. L.R.C. and R.R.D.; writing ? original draft, R.C.S. and M.S.G.; writing ? review & editing, R.C.S. C.W.V.K. A.A.D.R. P.J.R. C.J.W. and M.S.G.; funding acquisition, R.C.S. and M.S.G.; resources, R.C.S. and M.S.G.; supervision, R.C.S. and M.S.G. M.S.G. is a consultant for Maze Therapeutics, Enable Therapeutics, Glut1-Deficiency Syndrome Foundation, and Chelsea's Hope. M.S.G. R.C.S. C.W.V.K. and R.C.B. are founders of Atterogen, LLC.

Publisher Copyright:
© 2021 Elsevier Inc.

Keywords

Lafora disease
MALDI imaging
N-linked glycosylation
antibody-enzyme therapy
brain metabolism
childhood dementia
glucosamine
glycogen metabolism
glycogen storage disease
polyglucosan body

ASJC Scopus subject areas

Physiology
Molecular Biology
Cell Biology

UN SDGs

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

Access to Document

10.1016/j.cmet.2021.05.003

Cite this

Sun, R. C., Young, L. E. A., Bruntz, R. C., Markussen, K. H., Zhou, Z., Conroy, L. R., Hawkinson, T. R., Clarke, H. A., Stanback, A. E., Macedo, J. K. A., Emanuelle, S., Brewer, M. K., Rondon, A. L., Mestas, A., Sanders, W. C., Mahalingan, K. K., Tang, B., Chikwana, V. M., Segvich, D. M., ... Gentry, M. S. (2021). Brain glycogen serves as a critical glucosamine cache required for protein glycosylation. Cell Metabolism, 33(7), 1404-1417.e9. https://doi.org/10.1016/j.cmet.2021.05.003

@article{f5494e29ac5546318646ce514230592c,

title = "Brain glycogen serves as a critical glucosamine cache required for protein glycosylation",

abstract = "Glycosylation defects are a hallmark of many nervous system diseases. However, the molecular and metabolic basis for this pathology is not fully understood. In this study, we found that N-linked protein glycosylation in the brain is metabolically channeled to glucosamine metabolism through glycogenolysis. We discovered that glucosamine is an abundant constituent of brain glycogen, which functions as a glucosamine reservoir for multiple glycoconjugates. We demonstrated the enzymatic incorporation of glucosamine into glycogen by glycogen synthase, and the release by glycogen phosphorylase by biochemical and structural methodologies, in primary astrocytes, and in vivo by isotopic tracing and mass spectrometry. Using two mouse models of glycogen storage diseases, we showed that disruption of brain glycogen metabolism causes global decreases in free pools of UDP-N-acetylglucosamine and N-linked protein glycosylation. These findings revealed fundamental biological roles of brain glycogen in protein glycosylation with direct relevance to multiple human diseases of the central nervous system.",

keywords = "Lafora disease, MALDI imaging, N-linked glycosylation, antibody-enzyme therapy, brain metabolism, childhood dementia, glucosamine, glycogen metabolism, glycogen storage disease, polyglucosan body",

author = "Sun, {Ramon C.} and Young, {Lyndsay E.A.} and Bruntz, {Ronald C.} and Markussen, {Kia H.} and Zhengqiu Zhou and Conroy, {Lindsey R.} and Hawkinson, {Tara R.} and Clarke, {Harrison A.} and Stanback, {Alexandra E.} and Macedo, {Jessica K.A.} and Shane Emanuelle and Brewer, {M. Kathryn} and Rondon, {Alberto L.} and Annette Mestas and Sanders, {William C.} and Mahalingan, {Krishna K.} and Buyun Tang and Chikwana, {Vimbai M.} and Segvich, {Dyann M.} and Contreras, {Christopher J.} and Allenger, {Elizabeth J.} and Brainson, {Christine F.} and Johnson, {Lance A.} and Taylor, {Richard E.} and Armstrong, {Dustin D.} and Robert Shaffer and Waechter, {Charles J.} and {Vander Kooi}, {Craig W.} and DePaoli-Roach, {Anna A.} and Roach, {Peter J.} and Hurley, {Thomas D.} and Drake, {Richard R.} and Gentry, {Matthew S.}",

note = "Funding Information: We would like to thank Dr. Yokota for providing the anti-PGB antibody, Vander Kooi and Gentry lab members for vigorous discussions regarding the work, Mrs. Dana Napier for performing immunohistochemistry on tissue slices, and the Markey Cancer Center. The authors thank Nancy R. Gough (BioSerendipity, LLC) for editorial support. This study was supported by National Institute of Health (NIH) grants R35 NS116824 and P01 NS097197 to M.S.G.; NIH grant R01 AG066653 , St Baldrick{\textquoteright}s Career Development Award, V-Scholar Grant, and Rally Foundation Independent Investigator grant to R.C.S.; R01 DK27221 to P.J.R.; and NIH 5R01AG06255002 to L.A.J. J.K.A.M. and L.R.C. were supported by NIH / NCI training grant T32CA165990 . This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH -funded biospecimen Procurement & Translational Pathology Shared Resource Facility of the University of Kentucky Markey Cancer Center ( P30CA177558 ). We also acknowledge partial support of this work through the University of Notre Dame Reisenauer Family GSD Research Fund. Funding Information: We would like to thank Dr. Yokota for providing the anti-PGB antibody, Vander Kooi and Gentry lab members for vigorous discussions regarding the work, Mrs. Dana Napier for performing immunohistochemistry on tissue slices, and the Markey Cancer Center. The authors thank Nancy R. Gough (BioSerendipity, LLC) for editorial support. This study was supported by National Institute of Health (NIH) grants R35 NS116824 and P01 NS097197 to M.S.G.; NIH grant R01 AG066653, St Baldrick's Career Development Award, V-Scholar Grant, and Rally Foundation Independent Investigator grant to R.C.S.; R01 DK27221 to P.J.R.; and NIH 5R01AG06255002 to L.A.J. J.K.A.M. and L.R.C. were supported by NIH/NCI training grant T32CA165990. This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH-funded biospecimen Procurement & Translational Pathology Shared Resource Facility of the University of Kentucky Markey Cancer Center (P30CA177558). We also acknowledge partial support of this work through the University of Notre Dame Reisenauer Family GSD Research Fund. Conceptualization, R.C.S. and M.S.G.; methodology, R.C.S. M.S.G. and R.R.D.; investigation, R.C.S. L.E.A.Y. A.E.S. R.C.B. K.H.M. Z.Q.Z. J.K.A.M. S.E. M.K.B. A.L.R. W.C.S. K.K.M. B.T. V.M.C. D.M.S. C.J.C. E.J.A. C.F.B. L.A.J. R.E.T. C.W.V.K. C.W. A.A.D.R. P.J.R. T.D.H. L.R.C. and R.R.D.; writing ? original draft, R.C.S. and M.S.G.; writing ? review & editing, R.C.S. C.W.V.K. A.A.D.R. P.J.R. C.J.W. and M.S.G.; funding acquisition, R.C.S. and M.S.G.; resources, R.C.S. and M.S.G.; supervision, R.C.S. and M.S.G. M.S.G. is a consultant for Maze Therapeutics, Enable Therapeutics, Glut1-Deficiency Syndrome Foundation, and Chelsea's Hope. M.S.G. R.C.S. C.W.V.K. and R.C.B. are founders of Atterogen, LLC. Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",

year = "2021",

month = jul,

day = "6",

doi = "10.1016/j.cmet.2021.05.003",

language = "English",

volume = "33",

pages = "1404--1417.e9",

number = "7",

}

Sun, RC, Young, LEA, Bruntz, RC, Markussen, KH, Zhou, Z, Conroy, LR, Hawkinson, TR, Clarke, HA, Stanback, AE, Macedo, JKA, Emanuelle, S, Brewer, MK, Rondon, AL, Mestas, A, Sanders, WC, Mahalingan, KK, Tang, B, Chikwana, VM, Segvich, DM, Contreras, CJ, Allenger, EJ, Brainson, CF , Johnson, LA, Taylor, RE, Armstrong, DD, Shaffer, R, Waechter, CJ, Vander Kooi, CW, DePaoli-Roach, AA, Roach, PJ, Hurley, TD, Drake, RR & Gentry, MS 2021, 'Brain glycogen serves as a critical glucosamine cache required for protein glycosylation', Cell Metabolism, vol. 33, no. 7, pp. 1404-1417.e9. https://doi.org/10.1016/j.cmet.2021.05.003

TY - JOUR

T1 - Brain glycogen serves as a critical glucosamine cache required for protein glycosylation

AU - Sun, Ramon C.

AU - Young, Lyndsay E.A.

AU - Bruntz, Ronald C.

AU - Markussen, Kia H.

AU - Zhou, Zhengqiu

AU - Conroy, Lindsey R.

AU - Hawkinson, Tara R.

AU - Clarke, Harrison A.

AU - Stanback, Alexandra E.

AU - Macedo, Jessica K.A.

AU - Emanuelle, Shane

AU - Brewer, M. Kathryn

AU - Rondon, Alberto L.

AU - Mestas, Annette

AU - Sanders, William C.

AU - Mahalingan, Krishna K.

AU - Tang, Buyun

AU - Chikwana, Vimbai M.

AU - Segvich, Dyann M.

AU - Contreras, Christopher J.

AU - Allenger, Elizabeth J.

AU - Brainson, Christine F.

AU - Johnson, Lance A.

AU - Taylor, Richard E.

AU - Armstrong, Dustin D.

AU - Shaffer, Robert

AU - Waechter, Charles J.

AU - Vander Kooi, Craig W.

AU - DePaoli-Roach, Anna A.

AU - Roach, Peter J.

AU - Hurley, Thomas D.

AU - Drake, Richard R.

AU - Gentry, Matthew S.

N1 - Funding Information: We would like to thank Dr. Yokota for providing the anti-PGB antibody, Vander Kooi and Gentry lab members for vigorous discussions regarding the work, Mrs. Dana Napier for performing immunohistochemistry on tissue slices, and the Markey Cancer Center. The authors thank Nancy R. Gough (BioSerendipity, LLC) for editorial support. This study was supported by National Institute of Health (NIH) grants R35 NS116824 and P01 NS097197 to M.S.G.; NIH grant R01 AG066653 , St Baldrick’s Career Development Award, V-Scholar Grant, and Rally Foundation Independent Investigator grant to R.C.S.; R01 DK27221 to P.J.R.; and NIH 5R01AG06255002 to L.A.J. J.K.A.M. and L.R.C. were supported by NIH / NCI training grant T32CA165990 . This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH -funded biospecimen Procurement & Translational Pathology Shared Resource Facility of the University of Kentucky Markey Cancer Center ( P30CA177558 ). We also acknowledge partial support of this work through the University of Notre Dame Reisenauer Family GSD Research Fund. Funding Information: We would like to thank Dr. Yokota for providing the anti-PGB antibody, Vander Kooi and Gentry lab members for vigorous discussions regarding the work, Mrs. Dana Napier for performing immunohistochemistry on tissue slices, and the Markey Cancer Center. The authors thank Nancy R. Gough (BioSerendipity, LLC) for editorial support. This study was supported by National Institute of Health (NIH) grants R35 NS116824 and P01 NS097197 to M.S.G.; NIH grant R01 AG066653, St Baldrick's Career Development Award, V-Scholar Grant, and Rally Foundation Independent Investigator grant to R.C.S.; R01 DK27221 to P.J.R.; and NIH 5R01AG06255002 to L.A.J. J.K.A.M. and L.R.C. were supported by NIH/NCI training grant T32CA165990. This research was also supported by funding from the University of Kentucky Markey Cancer Center and the NIH-funded biospecimen Procurement & Translational Pathology Shared Resource Facility of the University of Kentucky Markey Cancer Center (P30CA177558). We also acknowledge partial support of this work through the University of Notre Dame Reisenauer Family GSD Research Fund. Conceptualization, R.C.S. and M.S.G.; methodology, R.C.S. M.S.G. and R.R.D.; investigation, R.C.S. L.E.A.Y. A.E.S. R.C.B. K.H.M. Z.Q.Z. J.K.A.M. S.E. M.K.B. A.L.R. W.C.S. K.K.M. B.T. V.M.C. D.M.S. C.J.C. E.J.A. C.F.B. L.A.J. R.E.T. C.W.V.K. C.W. A.A.D.R. P.J.R. T.D.H. L.R.C. and R.R.D.; writing ? original draft, R.C.S. and M.S.G.; writing ? review & editing, R.C.S. C.W.V.K. A.A.D.R. P.J.R. C.J.W. and M.S.G.; funding acquisition, R.C.S. and M.S.G.; resources, R.C.S. and M.S.G.; supervision, R.C.S. and M.S.G. M.S.G. is a consultant for Maze Therapeutics, Enable Therapeutics, Glut1-Deficiency Syndrome Foundation, and Chelsea's Hope. M.S.G. R.C.S. C.W.V.K. and R.C.B. are founders of Atterogen, LLC. Publisher Copyright: © 2021 Elsevier Inc.

PY - 2021/7/6

Y1 - 2021/7/6

N2 - Glycosylation defects are a hallmark of many nervous system diseases. However, the molecular and metabolic basis for this pathology is not fully understood. In this study, we found that N-linked protein glycosylation in the brain is metabolically channeled to glucosamine metabolism through glycogenolysis. We discovered that glucosamine is an abundant constituent of brain glycogen, which functions as a glucosamine reservoir for multiple glycoconjugates. We demonstrated the enzymatic incorporation of glucosamine into glycogen by glycogen synthase, and the release by glycogen phosphorylase by biochemical and structural methodologies, in primary astrocytes, and in vivo by isotopic tracing and mass spectrometry. Using two mouse models of glycogen storage diseases, we showed that disruption of brain glycogen metabolism causes global decreases in free pools of UDP-N-acetylglucosamine and N-linked protein glycosylation. These findings revealed fundamental biological roles of brain glycogen in protein glycosylation with direct relevance to multiple human diseases of the central nervous system.

AB - Glycosylation defects are a hallmark of many nervous system diseases. However, the molecular and metabolic basis for this pathology is not fully understood. In this study, we found that N-linked protein glycosylation in the brain is metabolically channeled to glucosamine metabolism through glycogenolysis. We discovered that glucosamine is an abundant constituent of brain glycogen, which functions as a glucosamine reservoir for multiple glycoconjugates. We demonstrated the enzymatic incorporation of glucosamine into glycogen by glycogen synthase, and the release by glycogen phosphorylase by biochemical and structural methodologies, in primary astrocytes, and in vivo by isotopic tracing and mass spectrometry. Using two mouse models of glycogen storage diseases, we showed that disruption of brain glycogen metabolism causes global decreases in free pools of UDP-N-acetylglucosamine and N-linked protein glycosylation. These findings revealed fundamental biological roles of brain glycogen in protein glycosylation with direct relevance to multiple human diseases of the central nervous system.

KW - Lafora disease

KW - MALDI imaging

KW - N-linked glycosylation

KW - antibody-enzyme therapy

KW - brain metabolism

KW - childhood dementia

KW - glucosamine

KW - glycogen metabolism

KW - glycogen storage disease

KW - polyglucosan body

UR - http://www.scopus.com/inward/record.url?scp=85109199166&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85109199166&partnerID=8YFLogxK

U2 - 10.1016/j.cmet.2021.05.003

DO - 10.1016/j.cmet.2021.05.003

M3 - Article

C2 - 34043942

AN - SCOPUS:85109199166

VL - 33

SP - 1404-1417.e9

IS - 7

ER -

Brain glycogen serves as a critical glucosamine cache required for protein glycosylation

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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