Interaction of a putative BH3 domain of clusterin with anti-apoptotic Bcl-2 family proteins as revealed by NMR spectroscopy

Dong Hwa Lee; Ji Hyang Ha; Yul Kim; Kwang Hee Bae; Jae Yong Park; Wan Sung Choi; Ho Sup Yoon; Sung Goo Park; Byoung Chul Park; Gwan Su Yi; Seung Wook Chi

doi:10.1016/j.bbrc.2011.04.054

Interaction of a putative BH3 domain of clusterin with anti-apoptotic Bcl-2 family proteins as revealed by NMR spectroscopy

Dong Hwa Lee, Ji Hyang Ha, Yul Kim, Kwang Hee Bae, Jae Yong Park, Wan Sung Choi, Ho Sup Yoon, Sung Goo Park, Byoung Chul Park, Gwan Su Yi, Seung Wook Chi

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

22 Citations (Scopus)

Abstract

Clusterin (CLU) is a multifunctional glycoprotein that is overexpressed in prostate and breast cancers. Although CLU is known to be involved in the regulation of apoptosis and cell survival, the precise molecular mechanism underlying the pro-apoptotic function of nuclear CLU (nCLU) remains unclear. In this study, we identified a conserved BH3 motif in C-terminal coiled coil (CC2) region of nCLU by sequence analysis and characterized the molecular interaction of the putative nCLU BH3 domain with anti-apoptotic Bcl-2 family proteins by nuclear magnetic resonance (NMR) spectroscopy. The chemical shift perturbation data demonstrated that the nCLU BH3 domain binds to pro-apoptotic BH3 peptide-binding grooves in both Bcl-X_L and Bcl-2. A structural model of the Bcl-X_L/nCLU BH3 peptide complex reveals that the binding mode is remarkably similar to those of other Bcl-X_L/BH3 peptide complexes. In addition, mutational analysis confirmed that Leu323 and Asp328 of nCLU BH3 domain, absolutely conserved in the BH3 motifs of BH3-only protein family, are critical for binding to Bcl-X_L. Taken altogether, our results suggest a molecular basis for the pro-apoptotic function of nCLU by elucidating the residue specific interactions of the BH3 motif in nCLU with anti-apoptotic Bcl-2 family proteins.

Original language	English
Pages (from-to)	541-547
Number of pages	7
Journal	Biochemical and biophysical research communications
Volume	408
Issue number	4
DOIs	https://doi.org/10.1016/j.bbrc.2011.04.054
Publication status	Published - 2011 May 20
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by Basic Science Research Program (No. 2009-0075665) and Mid-career Researcher Program through NRF grant funded by the MEST (No. 2008-0061624). This study was also supported by a grant from the National R&D Program for Cancer Control (0720130) and a grant of the Korean Health Technology R&D Project, Ministry for Health and Welfare, Republic of Korea (A100089). This study made use of the NMR facility at Korea Basic Science Institute, which is supported by Advanced Bio-MR technology Program of the MEST.

Keywords

Apoptosis
BH3 domain
Bcl-2 family protein
Clusterin
NMR

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

UN SDGs

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

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10.1016/j.bbrc.2011.04.054

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Lee, D. H., Ha, J. H., Kim, Y., Bae, K. H., Park, J. Y., Choi, W. S., Yoon, H. S., Park, S. G., Park, B. C., Yi, G. S., & Chi, S. W. (2011). Interaction of a putative BH3 domain of clusterin with anti-apoptotic Bcl-2 family proteins as revealed by NMR spectroscopy. Biochemical and biophysical research communications, 408(4), 541-547. https://doi.org/10.1016/j.bbrc.2011.04.054

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title = "Interaction of a putative BH3 domain of clusterin with anti-apoptotic Bcl-2 family proteins as revealed by NMR spectroscopy",

abstract = "Clusterin (CLU) is a multifunctional glycoprotein that is overexpressed in prostate and breast cancers. Although CLU is known to be involved in the regulation of apoptosis and cell survival, the precise molecular mechanism underlying the pro-apoptotic function of nuclear CLU (nCLU) remains unclear. In this study, we identified a conserved BH3 motif in C-terminal coiled coil (CC2) region of nCLU by sequence analysis and characterized the molecular interaction of the putative nCLU BH3 domain with anti-apoptotic Bcl-2 family proteins by nuclear magnetic resonance (NMR) spectroscopy. The chemical shift perturbation data demonstrated that the nCLU BH3 domain binds to pro-apoptotic BH3 peptide-binding grooves in both Bcl-XL and Bcl-2. A structural model of the Bcl-XL/nCLU BH3 peptide complex reveals that the binding mode is remarkably similar to those of other Bcl-XL/BH3 peptide complexes. In addition, mutational analysis confirmed that Leu323 and Asp328 of nCLU BH3 domain, absolutely conserved in the BH3 motifs of BH3-only protein family, are critical for binding to Bcl-XL. Taken altogether, our results suggest a molecular basis for the pro-apoptotic function of nCLU by elucidating the residue specific interactions of the BH3 motif in nCLU with anti-apoptotic Bcl-2 family proteins.",

keywords = "Apoptosis, BH3 domain, Bcl-2 family protein, Clusterin, NMR",

author = "Lee, {Dong Hwa} and Ha, {Ji Hyang} and Yul Kim and Bae, {Kwang Hee} and Park, {Jae Yong} and Choi, {Wan Sung} and Yoon, {Ho Sup} and Park, {Sung Goo} and Park, {Byoung Chul} and Yi, {Gwan Su} and Chi, {Seung Wook}",

note = "Funding Information: This work was supported by Basic Science Research Program (No. 2009-0075665) and Mid-career Researcher Program through NRF grant funded by the MEST (No. 2008-0061624). This study was also supported by a grant from the National R&D Program for Cancer Control (0720130) and a grant of the Korean Health Technology R&D Project, Ministry for Health and Welfare, Republic of Korea (A100089). This study made use of the NMR facility at Korea Basic Science Institute, which is supported by Advanced Bio-MR technology Program of the MEST.",

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AU - Lee, Dong Hwa

AU - Ha, Ji Hyang

AU - Kim, Yul

AU - Bae, Kwang Hee

AU - Park, Jae Yong

AU - Choi, Wan Sung

AU - Yoon, Ho Sup

AU - Park, Sung Goo

AU - Park, Byoung Chul

AU - Yi, Gwan Su

AU - Chi, Seung Wook

N1 - Funding Information: This work was supported by Basic Science Research Program (No. 2009-0075665) and Mid-career Researcher Program through NRF grant funded by the MEST (No. 2008-0061624). This study was also supported by a grant from the National R&D Program for Cancer Control (0720130) and a grant of the Korean Health Technology R&D Project, Ministry for Health and Welfare, Republic of Korea (A100089). This study made use of the NMR facility at Korea Basic Science Institute, which is supported by Advanced Bio-MR technology Program of the MEST.

PY - 2011/5/20

Y1 - 2011/5/20

N2 - Clusterin (CLU) is a multifunctional glycoprotein that is overexpressed in prostate and breast cancers. Although CLU is known to be involved in the regulation of apoptosis and cell survival, the precise molecular mechanism underlying the pro-apoptotic function of nuclear CLU (nCLU) remains unclear. In this study, we identified a conserved BH3 motif in C-terminal coiled coil (CC2) region of nCLU by sequence analysis and characterized the molecular interaction of the putative nCLU BH3 domain with anti-apoptotic Bcl-2 family proteins by nuclear magnetic resonance (NMR) spectroscopy. The chemical shift perturbation data demonstrated that the nCLU BH3 domain binds to pro-apoptotic BH3 peptide-binding grooves in both Bcl-XL and Bcl-2. A structural model of the Bcl-XL/nCLU BH3 peptide complex reveals that the binding mode is remarkably similar to those of other Bcl-XL/BH3 peptide complexes. In addition, mutational analysis confirmed that Leu323 and Asp328 of nCLU BH3 domain, absolutely conserved in the BH3 motifs of BH3-only protein family, are critical for binding to Bcl-XL. Taken altogether, our results suggest a molecular basis for the pro-apoptotic function of nCLU by elucidating the residue specific interactions of the BH3 motif in nCLU with anti-apoptotic Bcl-2 family proteins.

AB - Clusterin (CLU) is a multifunctional glycoprotein that is overexpressed in prostate and breast cancers. Although CLU is known to be involved in the regulation of apoptosis and cell survival, the precise molecular mechanism underlying the pro-apoptotic function of nuclear CLU (nCLU) remains unclear. In this study, we identified a conserved BH3 motif in C-terminal coiled coil (CC2) region of nCLU by sequence analysis and characterized the molecular interaction of the putative nCLU BH3 domain with anti-apoptotic Bcl-2 family proteins by nuclear magnetic resonance (NMR) spectroscopy. The chemical shift perturbation data demonstrated that the nCLU BH3 domain binds to pro-apoptotic BH3 peptide-binding grooves in both Bcl-XL and Bcl-2. A structural model of the Bcl-XL/nCLU BH3 peptide complex reveals that the binding mode is remarkably similar to those of other Bcl-XL/BH3 peptide complexes. In addition, mutational analysis confirmed that Leu323 and Asp328 of nCLU BH3 domain, absolutely conserved in the BH3 motifs of BH3-only protein family, are critical for binding to Bcl-XL. Taken altogether, our results suggest a molecular basis for the pro-apoptotic function of nCLU by elucidating the residue specific interactions of the BH3 motif in nCLU with anti-apoptotic Bcl-2 family proteins.

KW - Apoptosis

KW - BH3 domain

KW - Bcl-2 family protein

KW - Clusterin

KW - NMR

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SN - 0006-291X

VL - 408

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EP - 547

JO - Biochemical and biophysical research communications

JF - Biochemical and biophysical research communications

IS - 4

ER -

Interaction of a putative BH3 domain of clusterin with anti-apoptotic Bcl-2 family proteins as revealed by NMR spectroscopy

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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