skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes

Abstract

Lignin is a complex and irregular biopolymer of cross-linked phenylpropanoid units in plant secondary cell walls. Its biosynthesis requires three endoplasmic reticulum (ER)-resident cytochrome P450 monooxygenases, C4H, C3'H and F5H, to establish the structural characteristics of its monomeric precursors. Those P450 enzymes were reported to associate with each other or potentially with other soluble monolignol biosynthetic enzymes to form enzyme complex or metabolone. However, molecular basis governing such enzyme or pathway organization remains elusive. Here we show that Arabidopsis membrane steroid binding proteins (MSBPs) serve as scaffold physically organizing monolignol P450 monooxygenases thereby regulating lignin biosynthetic process. We find that although C4H, C3'H and F5H are in spatial proximity to each other on ER membrane in vivo, they do not appear to directly interact with each other. Instead, two MSBP proteins physically interact with all three P450 enzymes and, moreover, MSBPs themselves associate as homo- and heteromers on the ER membrane, thereby organizing P450 clusters. Down-regulation of MSBPs does not affect the transcription levels of monolignol biosynthetic P450 genes but substantially impairs the stability and activity of the MSBP-interacting P450 enzymes and, consequently, the lignin deposition, the accumulation of soluble phenolics in monolignol branch but not in flavonoid pathway. Furthermore,more » our study suggests that MSBP proteins are the essential structural component in ER membrane that physically organize and stabilize the monolignol biosynthetic P450 enzyme complex; thereby, specifically controlling phenylpropanoid-monolignol branch biosynthesis.« less

Authors:
ORCiD logo [1];  [1];  [2]; ORCiD logo [1]
  1. Brookhaven National Lab. (BNL), Upton, NY (United States)
  2. Stony Brook Univ., Stony Brook, NY (United States)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States); Energy Frontier Research Centers (EFRC) (United States). Center for Lignocellulose Structure and Formation (CLSF)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1462422
Report Number(s):
BNL-207910-2018-JAAM
Journal ID: ISSN 2055-0278
Grant/Contract Number:  
SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Nature Plants
Additional Journal Information:
Journal Volume: 4; Journal Issue: 5; Journal ID: ISSN 2055-0278
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Gou, Mingyue, Ran, Xiuzhi, Martin, Dwight W., and Liu, Chang -Jun. The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes. United States: N. p., 2018. Web. doi:10.1038/s41477-018-0142-9.
Gou, Mingyue, Ran, Xiuzhi, Martin, Dwight W., & Liu, Chang -Jun. The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes. United States. doi:10.1038/s41477-018-0142-9.
Gou, Mingyue, Ran, Xiuzhi, Martin, Dwight W., and Liu, Chang -Jun. Mon . "The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes". United States. doi:10.1038/s41477-018-0142-9. https://www.osti.gov/servlets/purl/1462422.
@article{osti_1462422,
title = {The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes},
author = {Gou, Mingyue and Ran, Xiuzhi and Martin, Dwight W. and Liu, Chang -Jun},
abstractNote = {Lignin is a complex and irregular biopolymer of cross-linked phenylpropanoid units in plant secondary cell walls. Its biosynthesis requires three endoplasmic reticulum (ER)-resident cytochrome P450 monooxygenases, C4H, C3'H and F5H, to establish the structural characteristics of its monomeric precursors. Those P450 enzymes were reported to associate with each other or potentially with other soluble monolignol biosynthetic enzymes to form enzyme complex or metabolone. However, molecular basis governing such enzyme or pathway organization remains elusive. Here we show that Arabidopsis membrane steroid binding proteins (MSBPs) serve as scaffold physically organizing monolignol P450 monooxygenases thereby regulating lignin biosynthetic process. We find that although C4H, C3'H and F5H are in spatial proximity to each other on ER membrane in vivo, they do not appear to directly interact with each other. Instead, two MSBP proteins physically interact with all three P450 enzymes and, moreover, MSBPs themselves associate as homo- and heteromers on the ER membrane, thereby organizing P450 clusters. Down-regulation of MSBPs does not affect the transcription levels of monolignol biosynthetic P450 genes but substantially impairs the stability and activity of the MSBP-interacting P450 enzymes and, consequently, the lignin deposition, the accumulation of soluble phenolics in monolignol branch but not in flavonoid pathway. Furthermore, our study suggests that MSBP proteins are the essential structural component in ER membrane that physically organize and stabilize the monolignol biosynthetic P450 enzyme complex; thereby, specifically controlling phenylpropanoid-monolignol branch biosynthesis.},
doi = {10.1038/s41477-018-0142-9},
journal = {Nature Plants},
number = 5,
volume = 4,
place = {United States},
year = {2018},
month = {4}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 17 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Lignin Biosynthesis
journal, June 2003


Dap1p, a Heme-Binding Protein That Regulates the Cytochrome P450 Protein Erg11p/Cyp51p in Saccharomyces cerevisiae
journal, February 2005


Spectroscopic and Biochemical Characterization of Heme Binding to Yeast Dap1p and Mouse PGRMC1p
journal, December 2005

  • Ghosh, Kaushik; Thompson, Alisha M.; Goldbeck, Robert A.
  • Biochemistry, Vol. 44, Issue 50
  • DOI: 10.1021/bi0511585

Cloning, Yeast Expression, and Characterization of the Coupling of Two Distantly Related Arabidopsis thaliana NADPH-Cytochrome P450 Reductases with P450 CYP73A5
journal, August 1997

  • Urban, Philippe; Mignotte, Claudia; Kazmaier, Michaël
  • Journal of Biological Chemistry, Vol. 272, Issue 31
  • DOI: 10.1074/jbc.272.31.19176

New GATEWAY vectors for High Throughput Analyses of Protein–Protein Interactions by Bimolecular Fluorescence Complementation
journal, September 2009

  • Gehl, Christian; Waadt, Rainer; Kudla, Jörg
  • Molecular Plant, Vol. 2, Issue 5
  • DOI: 10.1093/mp/ssp040

Regiospecific hydroxylation of isoflavones by cytochrome P450 81E enzymes from Medicago truncatula
journal, November 2003


Membrane protein complexes catalyze both 4- and 3-hydroxylation of cinnamic acid derivatives in monolignol biosynthesis
journal, December 2011

  • Chen, H. -C.; Li, Q.; Shuford, C. M.
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 52
  • DOI: 10.1073/pnas.1116416109

Arabidopsis Membrane Steroid Binding Protein 1 Is Involved in Inhibition of Cell Elongation
journal, December 2004


Evidence for enzyme complexes in the phenylpropanoid and flavonoid pathways
journal, September 1999


Metabolon formation and metabolic channeling in the biosynthesis of plant natural products
journal, June 2005

  • Jørgensen, Kirsten; Rasmussen, Anne Vinther; Morant, Marc
  • Current Opinion in Plant Biology, Vol. 8, Issue 3
  • DOI: 10.1016/j.pbi.2005.03.014

Membrane steroid-binding protein 1 (MSBP1) negatively regulates brassinosteroid signaling by enhancing the endocytosis of BAK1
journal, June 2009

  • Song, Li; Shi, Qiu-Ming; Yang, Xiao-Hua
  • Cell Research, Vol. 19, Issue 7
  • DOI: 10.1038/cr.2009.66

GATEWAY™ vectors for Agrobacterium-mediated plant transformation
journal, May 2002


The metabolon
journal, March 1985


Dap1/PGRMC1 Binds and Regulates Cytochrome P450 Enzymes
journal, February 2007


The Role of Protein-Protein and Protein-Membrane Interactions on P450 Function
journal, February 2016

  • Scott, E. E.; Wolf, C. R.; Otyepka, M.
  • Drug Metabolism and Disposition, Vol. 44, Issue 4
  • DOI: 10.1124/dmd.115.068569

The MYB107 Transcription Factor Positively Regulates Suberin Biosynthesis
journal, December 2016

  • Gou, Mingyue; Hou, Guichuan; Yang, Huijun
  • Plant Physiology, Vol. 173, Issue 2
  • DOI: 10.1104/pp.16.01614

Colocalization of l-Phenylalanine Ammonia-Lyase and Cinnamate 4-Hydroxylase for Metabolic Channeling in Phenylpropanoid Biosynthesis
journal, October 2004

  • Achnine, Lahoucine; Blancaflor, Elison B.; Rasmussen, Susanne
  • The Plant Cell, Vol. 16, Issue 11
  • DOI: 10.1105/tpc.104.024406

Phenylpropanoid Biosynthesis
journal, January 2010


K+ channel interactions detected by a genetic system optimized for systematic studies of membrane protein interactions
journal, August 2004

  • Obrdlik, P.; El-Bakkoury, M.; Hamacher, T.
  • Proceedings of the National Academy of Sciences, Vol. 101, Issue 33
  • DOI: 10.1073/pnas.0404467101

Characterization of the adrenal-specific antigen IZA (inner zone antigen) and its role in the steroidogenesis
journal, February 2004

  • Min, Li; Takemori, Hiroshi; Nonaka, Yasuki
  • Molecular and Cellular Endocrinology, Vol. 215, Issue 1-2
  • DOI: 10.1016/j.mce.2003.11.025

Arabidopsis Kelch Repeat F-Box Proteins Regulate Phenylpropanoid Biosynthesis via Controlling the Turnover of Phenylalanine Ammonia-Lyase
journal, December 2013


Split ubiquitin as a sensor of protein interactions in vivo.
journal, October 1994

  • Johnsson, N.; Varshavsky, A.
  • Proceedings of the National Academy of Sciences, Vol. 91, Issue 22
  • DOI: 10.1073/pnas.91.22.10340

Membrane Associated Progesterone Receptors: Promiscuous Proteins with Pleiotropic Functions – Focus on Interactions with Cytochromes P450
journal, March 2017


Emerging strategies of lignin engineering and degradation for cellulosic biofuel production
journal, April 2008


Molecular identification of adrenal inner zone antigen as a heme-binding protein: Molecular properties of adrenal inner zone antigen
journal, October 2005


Mutation of the Inducible ARABIDOPSIS THALIANA CYTOCHROME P450 REDUCTASE2 Alters Lignin Composition and Improves Saccharification
journal, October 2014

  • Sundin, Lisa; Vanholme, Ruben; Geerinck, Jan
  • Plant Physiology, Vol. 166, Issue 4
  • DOI: 10.1104/pp.114.245548

Linking genome and proteome by mass spectrometry: Large-scale identification of yeast proteins from two dimensional gels
journal, December 1996

  • Shevchenko, A.; Jensen, O. N.; Podtelejnikov, A. V.
  • Proceedings of the National Academy of Sciences, Vol. 93, Issue 25
  • DOI: 10.1073/pnas.93.25.14440

Protein–Protein and Protein–Membrane Associations in the Lignin Pathway
journal, November 2012

  • Bassard, Jean-Etienne; Richert, Ludovic; Geerinck, Jan
  • The Plant Cell, Vol. 24, Issue 11
  • DOI: 10.1105/tpc.112.102566

PGRMC1 (progesterone receptor membrane component 1): A targetable protein with multiple functions in steroid signaling, P450 activation and drug binding
journal, January 2009


Transgene-Mediated and Elicitor-Induced Perturbation of Metabolic Channeling at the Entry Point into the Phenylpropanoid Pathway
journal, August 1999


Spectroscopic and Mutagenesis Studies of Human PGRMC1
journal, February 2015

  • Kaluka, Daniel; Batabyal, Dipanwita; Chiang, Bing-Yu
  • Biochemistry, Vol. 54, Issue 8
  • DOI: 10.1021/bi501177e

Membrane Steroid Binding Protein 1 (MSBP1) Stimulates Tropism by Regulating Vesicle Trafficking and Auxin Redistribution
journal, November 2008

  • Yang, Xi; Song, Li; Xue, Hong-Wei
  • Molecular Plant, Vol. 1, Issue 6
  • DOI: 10.1093/mp/ssn071

A genetic system based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo
journal, April 1998

  • Stagljar, I.; Korostensky, C.; Johnsson, N.
  • Proceedings of the National Academy of Sciences, Vol. 95, Issue 9
  • DOI: 10.1073/pnas.95.9.5187

Heteromeric Complex Formation between CYP2E1 and CYP1A2:  Evidence for the Involvement of Electrostatic Interactions
journal, December 2006

  • Kelley, Rusty W.; Cheng, Dongmei; Backes, Wayne L.
  • Biochemistry, Vol. 45, Issue 51
  • DOI: 10.1021/bi061803n

Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation
journal, July 2007

  • Nakagawa, Tsuyoshi; Kurose, Takayuki; Hino, Takeshi
  • Journal of Bioscience and Bioengineering, Vol. 104, Issue 1
  • DOI: 10.1263/jbb.104.34

Bimolecular Fluorescence Complementation Analysis of Cytochrome p450 2c2, 2e1, and Nadph-Cytochrome p450 Reductase Molecular Interactions in Living Cells
journal, June 2005

  • Ozalp, Cengiz; Szczesna-Skorupa, Elzbieta; Kemper, Byron
  • Drug Metabolism and Disposition, Vol. 33, Issue 9
  • DOI: 10.1124/dmd.105.005538

Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance
journal, March 2016

  • Kabe, Yasuaki; Nakane, Takanori; Koike, Ikko
  • Nature Communications, Vol. 7, Issue 1
  • DOI: 10.1038/ncomms11030

Tobacco Transcription Factor WRKY1 Is Phosphorylated by the MAP Kinase SIPK and Mediates HR-Like Cell Death in Tobacco
journal, October 2005

  • Menke, Frank L. H.; Kang, Hong-Gu; Chen, Zhixiang
  • Molecular Plant-Microbe Interactions, Vol. 18, Issue 10
  • DOI: 10.1094/MPMI-18-1027

Twin anchors of the soybean isoflavonoid metabolon: evidence for tethering of the complex to the endoplasmic reticulum by IFS and C4H
journal, March 2016

  • Dastmalchi, Mehran; Bernards, Mark A.; Dhaubhadel, Sangeeta
  • The Plant Journal, Vol. 85, Issue 6
  • DOI: 10.1111/tpj.13137

Organization of multiple cytochrome P450s with NADPH-cytochrome P450 reductase in membranes
journal, May 2003


Cloning, Functional Expression, and Subcellular Localization of Multiple NADPH-Cytochrome P450 Reductases from Hybrid Poplar
journal, November 2002

  • Ro, Dae-Kyun; Ehlting, Jürgen; Douglas, Carl J.
  • Plant Physiology, Vol. 130, Issue 4
  • DOI: 10.1104/pp.008011

An Endoplasmic Reticulum-Derived Structure That Is Induced under Stress Conditions in Arabidopsis
journal, December 2002

  • Matsushima, Ryo; Hayashi, Yasuko; Kondo, Maki
  • Plant Physiology, Vol. 130, Issue 4
  • DOI: 10.1104/pp.009464

A Novel Role for Progesterone Receptor Membrane Component 1 (PGRMC1): A Partner and Regulator of Ferrochelatase
journal, September 2016


An Engineered Monolignol 4- O -Methyltransferase Depresses Lignin Biosynthesis and Confers Novel Metabolic Capability in Arabidopsis
journal, July 2012

  • Zhang, Kewei; Bhuiya, Mohammad-Wadud; Pazo, Jorge Rencoret
  • The Plant Cell, Vol. 24, Issue 7
  • DOI: 10.1105/tpc.112.101287

Split‐Ubiquitin System for Identifying Protein‐Protein Interactions in Membrane and Full‐Length Proteins
journal, October 2007


    Works referencing / citing this record:

    4-Coumarate 3-hydroxylase in the lignin biosynthesis pathway is a cytosolic ascorbate peroxidase
    journal, April 2019


    Comparative functional genomics analysis of cytochrome P450 gene superfamily in wheat and maize
    journal, March 2020


    Secondary cell wall biosynthesis
    journal, November 2018

    • Zhong, Ruiqin; Cui, Dongtao; Ye, Zheng‐Hua
    • New Phytologist, Vol. 221, Issue 4
    • DOI: 10.1111/nph.15537

    Lignocellulosic Biomass as Source for Lignin-Based Environmentally Benign Antioxidants
    journal, October 2018


    Glucosinolate and phenylpropanoid biosynthesis are linked by proteasome‐dependent degradation of PAL
    journal, September 2019

    • Kim, Jeong Im; Zhang, Xuebin; Pascuzzi, Pete E.
    • New Phytologist, Vol. 225, Issue 1
    • DOI: 10.1111/nph.16108

    Metabolons in plant primary and secondary metabolism
    journal, July 2019


    4-Coumarate 3-hydroxylase in the lignin biosynthesis pathway is a cytosolic ascorbate peroxidase
    journal, April 2019


    Proanthocyanidin subunit composition determined by functionally diverged dioxygenases
    journal, November 2018


    Glycoside‐specific glycosyltransferases catalyze regio‐selective sequential glucosylations for a sesame lignan, sesaminol triglucoside
    journal, December 2019

    • Ono, Eiichiro; Waki, Toshiyuki; Oikawa, Daiki
    • The Plant Journal, Vol. 101, Issue 5
    • DOI: 10.1111/tpj.14586

    Enzyme-Enzyme Interactions in Monolignol Biosynthesis
    journal, January 2019


    Dynamic metabolic solutions to the sessile life style of plants
    journal, January 2018

    • Knudsen, Camilla; Gallage, Nethaji Janeshawari; Hansen, Cecilie Cetti
    • Natural Product Reports, Vol. 35, Issue 11
    • DOI: 10.1039/c8np00037a

    Passive membrane transport of lignin-related compounds
    journal, October 2019

    • Vermaas, Josh V.; Dixon, Richard A.; Chen, Fang
    • Proceedings of the National Academy of Sciences, Vol. 116, Issue 46
    • DOI: 10.1073/pnas.1904643116

    Formation of Flavonoid Metabolons: Functional Significance of Protein-Protein Interactions and Impact on Flavonoid Chemodiversity
    journal, July 2019

    • Nakayama, Toru; Takahashi, Seiji; Waki, Toshiyuki
    • Frontiers in Plant Science, Vol. 10
    • DOI: 10.3389/fpls.2019.00821

    Lignin biosynthesis: old roads revisited and new roads explored
    journal, December 2019


    Enhanced Enzyme Activity through Scaffolding on Customizable Self‐Assembling Protein Filaments
    journal, March 2019


    The lignin toolbox of the model grass Setaria viridis
    journal, June 2019

    • Ferreira, Sávio Siqueira; Simões, Marcella Siqueira; Carvalho, Gabriel Garon
    • Plant Molecular Biology, Vol. 101, Issue 3
    • DOI: 10.1007/s11103-019-00897-9

    Lignin-Derived Biomaterials for Drug Release and Tissue Engineering
    journal, July 2018


    Formation of Flavonoid Metabolons: Functional Significance of Protein-Protein Interactions and Impact on Flavonoid Chemodiversity
    journal, July 2019

    • Nakayama, Toru; Takahashi, Seiji; Waki, Toshiyuki
    • Frontiers in Plant Science, Vol. 10
    • DOI: 10.3389/fpls.2019.00821

    Lignin-Derived Biomaterials for Drug Release and Tissue Engineering
    journal, July 2018


    The lignin toolbox of the model grass Setaria viridis
    journal, June 2019

    • Ferreira, Sávio Siqueira; Simões, Marcella Siqueira; Carvalho, Gabriel Garon
    • Plant Molecular Biology, Vol. 101, Issue 3
    • DOI: 10.1007/s11103-019-00897-9