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Esben Meldgaard Høgh Quistgaard
Structural and biochemical characterization of human PR70 in isolation and in complex with the scaffolding subunit of protein phosphatase 2A
Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review
DOI
- https://doi.org/10.1371/journal.pone.0101846
Final published version
- Rebecca Dovega, Department of Medical Biochemistry and Biophysics, Division of Physiological Chemistry II, Karolinska Institutet, Stockholm, Sweden. ,
- Susan Tsutakawa, Life Science Division, Lawrence Berkeley National Lab (LBNL), Berkeley, California, United States of America. ,
- Esben M Quistgaard
- Madhanagopal Anandapadamanaban, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden; Department of Physics, Chemistry and Biology, Linköping University, Linköping, Sweden. ,
- Christian Löw, Department of Medical Biochemistry and Biophysics, Division of Physiological Chemistry II, Karolinska Institutet, Stockholm, Sweden. ,
- Pär Nordlund, School of Biological Sciences, Nanyang Technological University, 637551, Singapore; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Protein Phosphatase 2A (PP2A) is a major Ser/Thr phosphatase involved in the regulation of various cellular processes. PP2A assembles into diverse trimeric holoenzymes, which consist of a scaffolding (A) subunit, a catalytic (C) subunit and various regulatory (B) subunits. Here we report a 2.0 Å crystal structure of the free B''/PR70 subunit and a SAXS model of an A/PR70 complex. The crystal structure of B''/PR70 reveals a two domain elongated structure with two Ca2+ binding EF-hands. Furthermore, we have characterized the interaction of both binding partner and their calcium dependency using biophysical techniques. Ca2+ biophysical studies with Circular Dichroism showed that the two EF-hands display different affinities to Ca2+. In the absence of the catalytic C-subunit, the scaffolding A-subunit remains highly mobile and flexible even in the presence of the B''/PR70 subunit as judged by SAXS. Isothermal Titration Calorimetry studies and SAXS data support that PR70 and the A-subunit have high affinity to each other. This study provides additional knowledge about the structural basis for the function of B'' containing holoenzymes.
| Original language | English |
|---|---|
| Journal | P L o S One |
| Volume | 9 |
| Issue | 7 |
| Pages (from-to) | e101846 |
| ISSN | 1932-6203 |
| DOIs | |
| Publication status | Published - 2014 |
| Externally published | Yes |
- Binding Sites, Calcium, Circular Dichroism, Crystallography, X-Ray, Holoenzymes, Humans, Models, Molecular, Protein Multimerization, Protein Phosphatase 2, Protein Structure, Secondary, Scattering, Small Angle, X-Ray Diffraction, Journal Article, Research Support, Non-U.S. Gov't
Research areas
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ID: 118858892