Structural Study of the HD-PTP Bro1 Domain in a Complex with the Core Region of STAM2, a Subunit of ESCRT-0

Juhyeon Lee; Kyoung-Jin Oh; Dasom Lee; Bo Yeon Kim; Joon Sig Choi; Bonsu Ku; Seung Jun Kim

doi:10.1371/journal.pone.0149113

Structural Study of the HD-PTP Bro1 Domain in a Complex with the Core Region of STAM2, a Subunit of ESCRT-0

PLoS One. 2016 Feb 11;11(2):e0149113. doi: 10.1371/journal.pone.0149113. eCollection 2016.

Authors

Juhyeon Lee^{1

2}, Kyoung-Jin Oh³, Dasom Lee³, Bo Yeon Kim⁴, Joon Sig Choi², Bonsu Ku¹, Seung Jun Kim¹

Affiliations

¹ Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
² Department of Biochemistry, Chungnam National University, Daejeon, Korea.
³ Research Center for Metabolic Regulation, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea.
⁴ Incurable Diseases Therapeutics Research Center, World Class Institute, Korea Research Institute of Bioscience and Biotechnology, Ochang, Cheongwon, Korea.

Abstract

EGFR is a key player in cell proliferation and survival signaling, and its sorting into MVBs for eventual lysosomal degradation is controlled by the coordination of multiple ESCRT complexes on the endosomal membrane. HD-PTP is a cytosolic protein tyrosine phosphatase, and is associated with EGFR trafficking by interacting with the ESCRT-0 protein STAM2 and the ESCRT-III protein CHMP4B via its N-terminal Bro1 domain. Intriguingly, the homologous domain of two other human Bro1 domain-containing proteins, Alix and Brox, binds CHMP4B but not STAM2, despite their high structural similarity. To elucidate this binding specificity, we determined the complex structure of the HD-PTP Bro1 domain bound to the STAM2 core region. STAM2 binds to the hydrophobic concave pocket of the HD-PTP Bro1 domain, as CHMP4B does to the pocket of Alix, Brox, or HD-PTP but in the opposite direction. Critically, Thr145 of HD-PTP, corresponding to Lys151 of Alix and Arg145 of Brox, is revealed to be a determinant residue enabling this protein to bind STAM2, as the Alix- or Brox-mimicking mutations of this residue blocks the intermolecular interaction. This work therefore provides the structural basis for how HD-PTP recognizes the ESCRT-0 component to control EGFR sorting.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptor Proteins, Signal Transducing / chemistry*
Calcium-Binding Proteins / chemistry
Calorimetry
Cell Cycle Proteins / chemistry
Cell Proliferation
Cell Survival
Crystallography, X-Ray
Endosomal Sorting Complexes Required for Transport / chemistry*
Endosomes / metabolism
ErbB Receptors / chemistry
Humans
Hydrophobic and Hydrophilic Interactions
Immunoblotting
Immunoprecipitation
Multivesicular Bodies / metabolism
Mutation
Protein Binding
Protein Structure, Secondary
Protein Structure, Tertiary
Protein Tyrosine Phosphatases, Non-Receptor / chemistry*
Recombinant Proteins / chemistry
Signal Transduction

Substances

Adaptor Proteins, Signal Transducing
BROX protein, human
CHMP4B protein, human
Calcium-Binding Proteins
Cell Cycle Proteins
Endosomal Sorting Complexes Required for Transport
PDCD6IP protein, human
Recombinant Proteins
STAM2 protein, human
EGFR protein, human
ErbB Receptors
PTPN23 protein, human
Protein Tyrosine Phosphatases, Non-Receptor

Grants and funding

This work was supported by grants from the National Research Foundation of Korea (2011-0030027 and 2015M3A9B5030308; to SJK) and the World Class Institute program (WCI 2009-002; to BYK) funded by the Ministry of Science, ICT and Future Planning. This work was also supported by the Korea Research Institute of Bioscience and Biotechnology Research Initiative Program for Creative Research (to BK). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.