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Article

Structural Analysis of the Complex between Penta-EF-Hand ALG-2 Protein and Sec31A Peptide Reveals a Novel Target Recognition Mechanism of ALG-2

1
Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan
2
Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-0801, Japan
3
Biomolecular Interaction Centre, School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch 8020, New Zealand
4
Department of Structural Biology, School of Medicine, Stanford University, Stanford, CA 94305-5126, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Charles A. Collyer
Int. J. Mol. Sci. 2015, 16(2), 3677-3699; https://doi.org/10.3390/ijms16023677
Received: 9 January 2015 / Accepted: 30 January 2015 / Published: 6 February 2015
(This article belongs to the Special Issue Protein Crystallography in Molecular Biology 2015)
ALG-2, a 22-kDa penta-EF-hand protein, is involved in cell death, signal transduction, membrane trafficking, etc., by interacting with various proteins in mammalian cells in a Ca2+-dependent manner. Most known ALG-2-interacting proteins contain proline-rich regions in which either PPYPXnYP (type 1 motif) or PXPGF (type 2 motif) is commonly found. Previous X-ray crystal structural analysis of the complex between ALG-2 and an ALIX peptide revealed that the peptide binds to the two hydrophobic pockets. In the present study, we resolved the crystal structure of the complex between ALG-2 and a peptide of Sec31A (outer shell component of coat complex II, COPII; containing the type 2 motif) and found that the peptide binds to the third hydrophobic pocket (Pocket 3). While amino acid substitution of Phe85, a Pocket 3 residue, with Ala abrogated the interaction with Sec31A, it did not affect the interaction with ALIX. On the other hand, amino acid substitution of Tyr180, a Pocket 1 residue, with Ala caused loss of binding to ALIX, but maintained binding to Sec31A. We conclude that ALG-2 recognizes two types of motifs at different hydrophobic surfaces. Furthermore, based on the results of serial mutational analysis of the ALG-2-binding sites in Sec31A, the type 2 motif was newly defined. View Full-Text
Keywords: adaptor protein; calcium-binding protein; COPII; crystal structure; EF-hand; motif; protein-protein interaction adaptor protein; calcium-binding protein; COPII; crystal structure; EF-hand; motif; protein-protein interaction
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MDPI and ACS Style

Takahashi, T.; Kojima, K.; Zhang, W.; Sasaki, K.; Ito, M.; Suzuki, H.; Kawasaki, M.; Wakatsuki, S.; Takahara, T.; Shibata, H.; Maki, M. Structural Analysis of the Complex between Penta-EF-Hand ALG-2 Protein and Sec31A Peptide Reveals a Novel Target Recognition Mechanism of ALG-2. Int. J. Mol. Sci. 2015, 16, 3677-3699. https://doi.org/10.3390/ijms16023677

AMA Style

Takahashi T, Kojima K, Zhang W, Sasaki K, Ito M, Suzuki H, Kawasaki M, Wakatsuki S, Takahara T, Shibata H, Maki M. Structural Analysis of the Complex between Penta-EF-Hand ALG-2 Protein and Sec31A Peptide Reveals a Novel Target Recognition Mechanism of ALG-2. International Journal of Molecular Sciences. 2015; 16(2):3677-3699. https://doi.org/10.3390/ijms16023677

Chicago/Turabian Style

Takahashi, Takeshi, Kyosuke Kojima, Wei Zhang, Kanae Sasaki, Masaru Ito, Hironori Suzuki, Masato Kawasaki, Soichi Wakatsuki, Terunao Takahara, Hideki Shibata, and Masatoshi Maki. 2015. "Structural Analysis of the Complex between Penta-EF-Hand ALG-2 Protein and Sec31A Peptide Reveals a Novel Target Recognition Mechanism of ALG-2" International Journal of Molecular Sciences 16, no. 2: 3677-3699. https://doi.org/10.3390/ijms16023677

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