Structural recognition mechanisms between human Src homology domain 3 (SH3) and ALG-2-interacting protein X (Alix)

Xiaoli Shi; Stephane Betzi; Adrien Lugari; Sandrine Opi; Audrey Restouin; Isabelle Parrot; Jean Martinez; Pascale Zimmermann; Patrick Lecine; Mingdong Huang; Stefan T. Arold; Yves Collette; Xavier Morelli

doi:10.1016/j.febslet.2012.05.017

Structural recognition mechanisms between human Src homology domain 3 (SH3) and ALG-2-interacting protein X (Alix)

Xiaoli Shi, Stephane Betzi, Adrien Lugari, Sandrine Opi, Audrey Restouin, Isabelle Parrot, Jean Martinez, Pascale Zimmermann, Patrick Lecine, Mingdong Huang, Stefan T. Arold^*, Yves Collette, Xavier Morelli

^*Corresponding author for this work

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

8 Scopus citations

Abstract

The functions of Src family kinases are tightly regulated through Src homology (SH) domain-mediated protein-protein interactions. We previously reported the biophysical characteristics of the apoptosis-linked gene 2-interacting protein X (Alix) in complex with the haemopoietic cell kinase (Hck) SH3 domain. In the current study, we have combined ITC, NMR, SAXS and molecular modeling to determine a 3D model of the complex. We demonstrate that Hck SH3 recognizes an extended linear proline-rich region of Alix. This particular binding mode enables Hck SH3 to sense a specific non-canonical residue situated in the SH3 RT-loop of the kinase. The resulting model helps clarify the mechanistic insights of Alix-Hck interaction. Structured summary of protein interactions: Hck physically interacts with SAM68 by two hybrid (View interaction) FynR96I physically interacts with Alix by two hybrid (View interaction) Hck binds to Alix by pull down (View interaction) Fyn physically interacts with SAM68 by two hybrid (View interaction) Hck and Alix bind by nuclear magnetic resonance (View interaction) FynR96I and Alix bind by isothermal titration calorimetry (View Interaction: 1, 2) FynR96I and Alix bind by nuclear magnetic resonance (View interaction) FynR96I binds to Alix by pull down (View interaction) Hck physically interacts with Alix by two hybrid (View interaction) FynR96I and Alix bind by x ray scattering (View interaction) Hck physically interacts with NEF by two hybrid (View interaction) FynR96I physically interacts with NEF by two hybrid (View interaction).

Original language	English (US)
Pages (from-to)	1759-1764
Number of pages	6
Journal	FEBS Letters
Volume	586
Issue number	13
DOIs	https://doi.org/10.1016/j.febslet.2012.05.017
State	Published - Jun 21 2012
Externally published	Yes

Keywords

Apoptosis-linked gene 2-interacting protein X (Alix)
NMR
Protein-protein interaction
SAXS
Src family kinase (SFK)
Src homology 3 domain (SH3 domain)

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/j.febslet.2012.05.017

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@article{45159b4dbf23400aa0b3cbb5b89a67c3,

title = "Structural recognition mechanisms between human Src homology domain 3 (SH3) and ALG-2-interacting protein X (Alix)",

abstract = "The functions of Src family kinases are tightly regulated through Src homology (SH) domain-mediated protein-protein interactions. We previously reported the biophysical characteristics of the apoptosis-linked gene 2-interacting protein X (Alix) in complex with the haemopoietic cell kinase (Hck) SH3 domain. In the current study, we have combined ITC, NMR, SAXS and molecular modeling to determine a 3D model of the complex. We demonstrate that Hck SH3 recognizes an extended linear proline-rich region of Alix. This particular binding mode enables Hck SH3 to sense a specific non-canonical residue situated in the SH3 RT-loop of the kinase. The resulting model helps clarify the mechanistic insights of Alix-Hck interaction. Structured summary of protein interactions: Hck physically interacts with SAM68 by two hybrid (View interaction) FynR96I physically interacts with Alix by two hybrid (View interaction) Hck binds to Alix by pull down (View interaction) Fyn physically interacts with SAM68 by two hybrid (View interaction) Hck and Alix bind by nuclear magnetic resonance (View interaction) FynR96I and Alix bind by isothermal titration calorimetry (View Interaction: 1, 2) FynR96I and Alix bind by nuclear magnetic resonance (View interaction) FynR96I binds to Alix by pull down (View interaction) Hck physically interacts with Alix by two hybrid (View interaction) FynR96I and Alix bind by x ray scattering (View interaction) Hck physically interacts with NEF by two hybrid (View interaction) FynR96I physically interacts with NEF by two hybrid (View interaction).",

keywords = "Apoptosis-linked gene 2-interacting protein X (Alix), NMR, Protein-protein interaction, SAXS, Src family kinase (SFK), Src homology 3 domain (SH3 domain)",

author = "Xiaoli Shi and Stephane Betzi and Adrien Lugari and Sandrine Opi and Audrey Restouin and Isabelle Parrot and Jean Martinez and Pascale Zimmermann and Patrick Lecine and Mingdong Huang and Arold, {Stefan T.} and Yves Collette and Xavier Morelli",

note = "Funding Information: This work was supported by the Centre National de la Recherche Scientifique (CNRS); the Institut National de la Sant{\'e} et de la Recherche M{\'e}dicale (INSERM); the Agence Nationale de Recherche sur le SIDA et les h{\'e}patites virales (ANRS); the Ministry of Science and Technology, China [Grant Nos. 2007CB914304; 2006AA02A313]; the National Natural Science Foundation of China (NSFC) [Grant Nos. 30800181; 30625011]; the Research Foundation Flanders (FWO); the concerted action programme of the Katholieke Universiteit Leuven; and the National Institutes of Health (MD Anderson{\textquoteright}s Cancer Center Support) [Grant No. CA016672]. S.O. and A.L. were fellows of the ANRS, and X.S. was a fellow of the {\textquoteleft}Ambassade de France en Chine{\textquoteright}. ",

year = "2012",

month = jun,

day = "21",

doi = "10.1016/j.febslet.2012.05.017",

language = "English (US)",

volume = "586",

pages = "1759--1764",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "Wiley",

number = "13",

}

TY - JOUR

T1 - Structural recognition mechanisms between human Src homology domain 3 (SH3) and ALG-2-interacting protein X (Alix)

AU - Shi, Xiaoli

AU - Betzi, Stephane

AU - Lugari, Adrien

AU - Opi, Sandrine

AU - Restouin, Audrey

AU - Parrot, Isabelle

AU - Martinez, Jean

AU - Zimmermann, Pascale

AU - Lecine, Patrick

AU - Huang, Mingdong

AU - Arold, Stefan T.

AU - Collette, Yves

AU - Morelli, Xavier

N1 - Funding Information: This work was supported by the Centre National de la Recherche Scientifique (CNRS); the Institut National de la Santé et de la Recherche Médicale (INSERM); the Agence Nationale de Recherche sur le SIDA et les hépatites virales (ANRS); the Ministry of Science and Technology, China [Grant Nos. 2007CB914304; 2006AA02A313]; the National Natural Science Foundation of China (NSFC) [Grant Nos. 30800181; 30625011]; the Research Foundation Flanders (FWO); the concerted action programme of the Katholieke Universiteit Leuven; and the National Institutes of Health (MD Anderson’s Cancer Center Support) [Grant No. CA016672]. S.O. and A.L. were fellows of the ANRS, and X.S. was a fellow of the ‘Ambassade de France en Chine’.

PY - 2012/6/21

Y1 - 2012/6/21

N2 - The functions of Src family kinases are tightly regulated through Src homology (SH) domain-mediated protein-protein interactions. We previously reported the biophysical characteristics of the apoptosis-linked gene 2-interacting protein X (Alix) in complex with the haemopoietic cell kinase (Hck) SH3 domain. In the current study, we have combined ITC, NMR, SAXS and molecular modeling to determine a 3D model of the complex. We demonstrate that Hck SH3 recognizes an extended linear proline-rich region of Alix. This particular binding mode enables Hck SH3 to sense a specific non-canonical residue situated in the SH3 RT-loop of the kinase. The resulting model helps clarify the mechanistic insights of Alix-Hck interaction. Structured summary of protein interactions: Hck physically interacts with SAM68 by two hybrid (View interaction) FynR96I physically interacts with Alix by two hybrid (View interaction) Hck binds to Alix by pull down (View interaction) Fyn physically interacts with SAM68 by two hybrid (View interaction) Hck and Alix bind by nuclear magnetic resonance (View interaction) FynR96I and Alix bind by isothermal titration calorimetry (View Interaction: 1, 2) FynR96I and Alix bind by nuclear magnetic resonance (View interaction) FynR96I binds to Alix by pull down (View interaction) Hck physically interacts with Alix by two hybrid (View interaction) FynR96I and Alix bind by x ray scattering (View interaction) Hck physically interacts with NEF by two hybrid (View interaction) FynR96I physically interacts with NEF by two hybrid (View interaction).

AB - The functions of Src family kinases are tightly regulated through Src homology (SH) domain-mediated protein-protein interactions. We previously reported the biophysical characteristics of the apoptosis-linked gene 2-interacting protein X (Alix) in complex with the haemopoietic cell kinase (Hck) SH3 domain. In the current study, we have combined ITC, NMR, SAXS and molecular modeling to determine a 3D model of the complex. We demonstrate that Hck SH3 recognizes an extended linear proline-rich region of Alix. This particular binding mode enables Hck SH3 to sense a specific non-canonical residue situated in the SH3 RT-loop of the kinase. The resulting model helps clarify the mechanistic insights of Alix-Hck interaction. Structured summary of protein interactions: Hck physically interacts with SAM68 by two hybrid (View interaction) FynR96I physically interacts with Alix by two hybrid (View interaction) Hck binds to Alix by pull down (View interaction) Fyn physically interacts with SAM68 by two hybrid (View interaction) Hck and Alix bind by nuclear magnetic resonance (View interaction) FynR96I and Alix bind by isothermal titration calorimetry (View Interaction: 1, 2) FynR96I and Alix bind by nuclear magnetic resonance (View interaction) FynR96I binds to Alix by pull down (View interaction) Hck physically interacts with Alix by two hybrid (View interaction) FynR96I and Alix bind by x ray scattering (View interaction) Hck physically interacts with NEF by two hybrid (View interaction) FynR96I physically interacts with NEF by two hybrid (View interaction).

KW - Apoptosis-linked gene 2-interacting protein X (Alix)

KW - NMR

KW - Protein-protein interaction

KW - SAXS

KW - Src family kinase (SFK)

KW - Src homology 3 domain (SH3 domain)

UR - http://www.scopus.com/inward/record.url?scp=84862501603&partnerID=8YFLogxK

U2 - 10.1016/j.febslet.2012.05.017

DO - 10.1016/j.febslet.2012.05.017

M3 - Article

C2 - 22641034

AN - SCOPUS:84862501603

SN - 0014-5793

VL - 586

SP - 1759

EP - 1764

JO - FEBS Letters

JF - FEBS Letters

IS - 13

ER -

Structural recognition mechanisms between human Src homology domain 3 (SH3) and ALG-2-interacting protein X (Alix)

Abstract

Keywords

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