Unique structural features of a BCL-2 family protein CED-9 and biophysical characterization of CED-9/EGL-1 interactions

J. S. Woo; J. S. Jung; N. C. Ha; J. Shin; K. H. Kim; W. Lee; B. H. Oh

doi:10.1038/sj.cdd.4401303

Unique structural features of a BCL-2 family protein CED-9 and biophysical characterization of CED-9/EGL-1 interactions

J. S. Woo, J. S. Jung, N. C. Ha, J. Shin, K. H. Kim, W. Lee, B. H. Oh

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

37 Citations (Scopus)

Abstract

The interactions between B-cell lymphoma 2 (BCL-2) family members are known to be mediated through the binding of the BH3 domain of a proapoptotic member to the BH3-binding groove of an antiapoptotic member. We determined the crystal structure of antiapoptotic CED-9, which reveals a unique C-terminal helix altering the common BH3-binding region. A coexpression system to produce CED-9 in complex with proapoptotic EGL-1 enabled us to show that the binding of EGL-1 to CED-9 is extremely stable, raising the melting temperature (T_m) of CED-9 by 25°C, and that the binding surface of CED-9 extends beyond the BH3-binding region and reaches the BH4 domain. Consistently, the T_m and a ¹H-¹⁵N correlation NMR spectrum of CED-9 in complex with EGL-1 are drastically different from those of CED-9 in complex with the EGL-1 BH3 peptide. The data suggest that the recognition between other BCL-2 family members may also involve much wider protein surfaces than is previously thought.

Original language	English
Pages (from-to)	1310-1319
Number of pages	10
Journal	Cell Death and Differentiation
Volume	10
Issue number	12
DOIs	https://doi.org/10.1038/sj.cdd.4401303
Publication status	Published - 2003 Dec
Externally published	Yes

Bibliographical note

Funding Information:
We thank Professors J Ahnn (KJIST) and J Lee (Yonsei University) for providing the C. elegance cDNA library. This study used the beamline 6B at the Pohang Accelerator Laboratory and was supported by Creative Research Initiatives (to B-HO) and by the National Research Laboratory program (M1-0203-00-0020) (to WL) of the Korean Ministry of Science and Technology. J-SW, J-SJ, and N-CH were supported by the Brain Korea 21 Project.

Keywords

Apoptosis
BCL-2 family
CED-9
EGL-1
Molecular recognition

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1038/sj.cdd.4401303

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title = "Unique structural features of a BCL-2 family protein CED-9 and biophysical characterization of CED-9/EGL-1 interactions",

abstract = "The interactions between B-cell lymphoma 2 (BCL-2) family members are known to be mediated through the binding of the BH3 domain of a proapoptotic member to the BH3-binding groove of an antiapoptotic member. We determined the crystal structure of antiapoptotic CED-9, which reveals a unique C-terminal helix altering the common BH3-binding region. A coexpression system to produce CED-9 in complex with proapoptotic EGL-1 enabled us to show that the binding of EGL-1 to CED-9 is extremely stable, raising the melting temperature (Tm) of CED-9 by 25°C, and that the binding surface of CED-9 extends beyond the BH3-binding region and reaches the BH4 domain. Consistently, the Tm and a 1H-15N correlation NMR spectrum of CED-9 in complex with EGL-1 are drastically different from those of CED-9 in complex with the EGL-1 BH3 peptide. The data suggest that the recognition between other BCL-2 family members may also involve much wider protein surfaces than is previously thought.",

keywords = "Apoptosis, BCL-2 family, CED-9, EGL-1, Molecular recognition",

author = "Woo, {J. S.} and Jung, {J. S.} and Ha, {N. C.} and J. Shin and Kim, {K. H.} and W. Lee and Oh, {B. H.}",

note = "Funding Information: We thank Professors J Ahnn (KJIST) and J Lee (Yonsei University) for providing the C. elegance cDNA library. This study used the beamline 6B at the Pohang Accelerator Laboratory and was supported by Creative Research Initiatives (to B-HO) and by the National Research Laboratory program (M1-0203-00-0020) (to WL) of the Korean Ministry of Science and Technology. J-SW, J-SJ, and N-CH were supported by the Brain Korea 21 Project.",

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AU - Shin, J.

AU - Kim, K. H.

AU - Lee, W.

AU - Oh, B. H.

N1 - Funding Information: We thank Professors J Ahnn (KJIST) and J Lee (Yonsei University) for providing the C. elegance cDNA library. This study used the beamline 6B at the Pohang Accelerator Laboratory and was supported by Creative Research Initiatives (to B-HO) and by the National Research Laboratory program (M1-0203-00-0020) (to WL) of the Korean Ministry of Science and Technology. J-SW, J-SJ, and N-CH were supported by the Brain Korea 21 Project.

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N2 - The interactions between B-cell lymphoma 2 (BCL-2) family members are known to be mediated through the binding of the BH3 domain of a proapoptotic member to the BH3-binding groove of an antiapoptotic member. We determined the crystal structure of antiapoptotic CED-9, which reveals a unique C-terminal helix altering the common BH3-binding region. A coexpression system to produce CED-9 in complex with proapoptotic EGL-1 enabled us to show that the binding of EGL-1 to CED-9 is extremely stable, raising the melting temperature (Tm) of CED-9 by 25°C, and that the binding surface of CED-9 extends beyond the BH3-binding region and reaches the BH4 domain. Consistently, the Tm and a 1H-15N correlation NMR spectrum of CED-9 in complex with EGL-1 are drastically different from those of CED-9 in complex with the EGL-1 BH3 peptide. The data suggest that the recognition between other BCL-2 family members may also involve much wider protein surfaces than is previously thought.

AB - The interactions between B-cell lymphoma 2 (BCL-2) family members are known to be mediated through the binding of the BH3 domain of a proapoptotic member to the BH3-binding groove of an antiapoptotic member. We determined the crystal structure of antiapoptotic CED-9, which reveals a unique C-terminal helix altering the common BH3-binding region. A coexpression system to produce CED-9 in complex with proapoptotic EGL-1 enabled us to show that the binding of EGL-1 to CED-9 is extremely stable, raising the melting temperature (Tm) of CED-9 by 25°C, and that the binding surface of CED-9 extends beyond the BH3-binding region and reaches the BH4 domain. Consistently, the Tm and a 1H-15N correlation NMR spectrum of CED-9 in complex with EGL-1 are drastically different from those of CED-9 in complex with the EGL-1 BH3 peptide. The data suggest that the recognition between other BCL-2 family members may also involve much wider protein surfaces than is previously thought.

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Unique structural features of a BCL-2 family protein CED-9 and biophysical characterization of CED-9/EGL-1 interactions

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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