Pro-metastatic collagen lysyl hydroxylase dimer assemblies stabilized by Fe2+-binding

Hou Fu Guo; Chi Lin Tsai; Masahiko Terajima; Xiaochao Tan; Priyam Banerjee; Mitchell D. Miller; Xin Liu; Jiang Yu; Jovita Byemerwa; Sarah Alvarado; Tamer S. Kaoud; Kevin N. Dalby; Neus Bota-Rabassedas; Yulong Chen; Mitsuo Yamauchi; John A. Tainer; George N. Phillips; Jonathan M. Kurie

doi:10.1038/s41467-018-02859-z

Pro-metastatic collagen lysyl hydroxylase dimer assemblies stabilized by Fe²⁺-binding

Hou Fu Guo, Chi Lin Tsai, Masahiko Terajima, Xiaochao Tan, Priyam Banerjee, Mitchell D. Miller, Xin Liu, Jiang Yu, Jovita Byemerwa, Sarah Alvarado, Tamer S. Kaoud, Kevin N. Dalby, Neus Bota-Rabassedas, Yulong Chen, Mitsuo Yamauchi, John A. Tainer, George N. Phillips, Jonathan M. Kurie

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

43 Scopus citations

Abstract

Collagen lysyl hydroxylases (LH1-3) are Fe²⁺- and 2-oxoglutarate (2-OG)-dependent oxygenases that maintain extracellular matrix homeostasis. High LH2 levels cause stable collagen cross-link accumulations that promote fibrosis and cancer progression. However, developing LH antagonists will require structural insights. Here, we report a 2 Å crystal structure and X-ray scattering on dimer assemblies for the LH domain of L230 in Acanthamoeba polyphaga mimivirus. Loop residues in the double-stranded β-helix core generate a tail-to-tail dimer. A stabilizing hydrophobic leucine locks into an aromatic tyrosine-pocket on the opposite subunit. An active site triad coordinates Fe²⁺. The two active sites flank a deep surface cleft that suggest dimerization creates a collagen-binding site. Loss of Fe²⁺-binding disrupts the dimer. Dimer disruption and charge reversal in the cleft increase K _m and reduce LH activity. Ectopic L230 expression in tumors promotes collagen cross-linking and metastasis. These insights suggest inhibitor targets for fibrosis and cancer.

Original language	English
Article number	512
Journal	Nature Communications
Volume	9
Issue number	1
DOIs	https://doi.org/10.1038/s41467-018-02859-z
State	Published - Dec 1 2018

Bibliographical note

Publisher Copyright:
© 2018 The Author(s).

Funding

This work was supported in part by the National Institutes of Health grants R21AR060978 (M.Y.), R01CA105155 (J.M.K. and M.Y.), P50CA70907 (J.M.K.); Cancer Prevention and Research Institute of Texas (CPRIT) grant RP160652 (J.M.K.); The International Association for the Study of Lung Cancer Postdoctoral Fellowship (H.-F.G.); the Elza A. and Ina S. Freeman Professorship in Lung Cancer (J.M.K.); and The Welch Foundation F-1390 (K.N.D.); NIH GM123252 (K.N.D.); and CPRIT RP160657 (K.N.D.). We thank Kathryn Brunett and the SIBYLS beamline staff for collecting SAXS data. The SIBYLS beamline is supported through the Integrated Diffraction Analysis Technologies Program, which is supported by Department of Energy Office of Biological and Environmental Research, National Institute of Health project MINOS (R01GM105404), and a High-End Instrumentation Grant S10OD018483. We thank Andre Mueller from Wyatt Technology for MALS data analysis support.

Funders	Funder number
Office of Biological and Environmental Research of the Department of Energy
Elza A.
National Institutes of Health/National Institute of Environmental Health Sciences	R01GM105404, S10OD018483
National Institutes of Health (NIH)	R01CA105155, R21AR060978, CPRIT RP160657, GM123252, P50CA70907
National Institute of General Medical Sciences	T32GM008280
Welch Foundation
Cancer Prevention and Research Institute of Texas	RP160652
International Association for the Study of Lung Cancer

ASJC Scopus subject areas

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

UN SDGs

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

Access to Document

10.1038/s41467-018-02859-z

Cite this

Guo, H. F., Tsai, C. L., Terajima, M., Tan, X., Banerjee, P., Miller, M. D., Liu, X., Yu, J., Byemerwa, J., Alvarado, S., Kaoud, T. S., Dalby, K. N., Bota-Rabassedas, N., Chen, Y., Yamauchi, M., Tainer, J. A., Phillips, G. N., & Kurie, J. M. (2018). Pro-metastatic collagen lysyl hydroxylase dimer assemblies stabilized by Fe²⁺-binding. Nature Communications, 9(1), Article 512. https://doi.org/10.1038/s41467-018-02859-z

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title = "Pro-metastatic collagen lysyl hydroxylase dimer assemblies stabilized by Fe2+-binding",

abstract = "Collagen lysyl hydroxylases (LH1-3) are Fe2+- and 2-oxoglutarate (2-OG)-dependent oxygenases that maintain extracellular matrix homeostasis. High LH2 levels cause stable collagen cross-link accumulations that promote fibrosis and cancer progression. However, developing LH antagonists will require structural insights. Here, we report a 2 {\AA} crystal structure and X-ray scattering on dimer assemblies for the LH domain of L230 in Acanthamoeba polyphaga mimivirus. Loop residues in the double-stranded β-helix core generate a tail-to-tail dimer. A stabilizing hydrophobic leucine locks into an aromatic tyrosine-pocket on the opposite subunit. An active site triad coordinates Fe2+. The two active sites flank a deep surface cleft that suggest dimerization creates a collagen-binding site. Loss of Fe2+-binding disrupts the dimer. Dimer disruption and charge reversal in the cleft increase K m and reduce LH activity. Ectopic L230 expression in tumors promotes collagen cross-linking and metastasis. These insights suggest inhibitor targets for fibrosis and cancer.",

author = "Guo, \{Hou Fu\} and Tsai, \{Chi Lin\} and Masahiko Terajima and Xiaochao Tan and Priyam Banerjee and Miller, \{Mitchell D.\} and Xin Liu and Jiang Yu and Jovita Byemerwa and Sarah Alvarado and Kaoud, \{Tamer S.\} and Dalby, \{Kevin N.\} and Neus Bota-Rabassedas and Yulong Chen and Mitsuo Yamauchi and Tainer, \{John A.\} and Phillips, \{George N.\} and Kurie, \{Jonathan M.\}",

note = "Publisher Copyright: {\textcopyright} 2018 The Author(s).",

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doi = "10.1038/s41467-018-02859-z",

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Guo, HF, Tsai, CL, Terajima, M, Tan, X, Banerjee, P, Miller, MD, Liu, X, Yu, J, Byemerwa, J, Alvarado, S, Kaoud, TS, Dalby, KN, Bota-Rabassedas, N, Chen, Y, Yamauchi, M, Tainer, JA, Phillips, GN & Kurie, JM 2018, 'Pro-metastatic collagen lysyl hydroxylase dimer assemblies stabilized by Fe²⁺-binding', Nature Communications, vol. 9, no. 1, 512. https://doi.org/10.1038/s41467-018-02859-z

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AU - Guo, Hou Fu

AU - Tsai, Chi Lin

AU - Terajima, Masahiko

AU - Tan, Xiaochao

AU - Banerjee, Priyam

AU - Miller, Mitchell D.

AU - Liu, Xin

AU - Yu, Jiang

AU - Byemerwa, Jovita

AU - Alvarado, Sarah

AU - Kaoud, Tamer S.

AU - Dalby, Kevin N.

AU - Bota-Rabassedas, Neus

AU - Chen, Yulong

AU - Yamauchi, Mitsuo

AU - Tainer, John A.

AU - Phillips, George N.

AU - Kurie, Jonathan M.

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Collagen lysyl hydroxylases (LH1-3) are Fe2+- and 2-oxoglutarate (2-OG)-dependent oxygenases that maintain extracellular matrix homeostasis. High LH2 levels cause stable collagen cross-link accumulations that promote fibrosis and cancer progression. However, developing LH antagonists will require structural insights. Here, we report a 2 Å crystal structure and X-ray scattering on dimer assemblies for the LH domain of L230 in Acanthamoeba polyphaga mimivirus. Loop residues in the double-stranded β-helix core generate a tail-to-tail dimer. A stabilizing hydrophobic leucine locks into an aromatic tyrosine-pocket on the opposite subunit. An active site triad coordinates Fe2+. The two active sites flank a deep surface cleft that suggest dimerization creates a collagen-binding site. Loss of Fe2+-binding disrupts the dimer. Dimer disruption and charge reversal in the cleft increase K m and reduce LH activity. Ectopic L230 expression in tumors promotes collagen cross-linking and metastasis. These insights suggest inhibitor targets for fibrosis and cancer.

AB - Collagen lysyl hydroxylases (LH1-3) are Fe2+- and 2-oxoglutarate (2-OG)-dependent oxygenases that maintain extracellular matrix homeostasis. High LH2 levels cause stable collagen cross-link accumulations that promote fibrosis and cancer progression. However, developing LH antagonists will require structural insights. Here, we report a 2 Å crystal structure and X-ray scattering on dimer assemblies for the LH domain of L230 in Acanthamoeba polyphaga mimivirus. Loop residues in the double-stranded β-helix core generate a tail-to-tail dimer. A stabilizing hydrophobic leucine locks into an aromatic tyrosine-pocket on the opposite subunit. An active site triad coordinates Fe2+. The two active sites flank a deep surface cleft that suggest dimerization creates a collagen-binding site. Loss of Fe2+-binding disrupts the dimer. Dimer disruption and charge reversal in the cleft increase K m and reduce LH activity. Ectopic L230 expression in tumors promotes collagen cross-linking and metastasis. These insights suggest inhibitor targets for fibrosis and cancer.

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