Studying Conformational Changes of the Yersinia Type-III-Secretion Effector YopO in Solution by Integrative Structural Biology

Martin F Peter; Anne T Tuukkanen; Caspar A Heubach; Alexander Selsam; Fraser G Duthie; Dmitri I Svergun; Olav Schiemann; Gregor Hagelueken

doi:10.1016/j.str.2019.06.007

Studying Conformational Changes of the Yersinia Type-III-Secretion Effector YopO in Solution by Integrative Structural Biology

Structure. 2019 Sep 3;27(9):1416-1426.e3. doi: 10.1016/j.str.2019.06.007. Epub 2019 Jul 11.

Authors

Martin F Peter¹, Anne T Tuukkanen², Caspar A Heubach¹, Alexander Selsam¹, Fraser G Duthie¹, Dmitri I Svergun², Olav Schiemann¹, Gregor Hagelueken³

Affiliations

¹ Institute for Physical and Theoretical Chemistry, University of Bonn, 53115 Bonn, Germany.
² European Molecular Biology Laboratory, Hamburg Unit, 22607 Hamburg, Germany.
³ Institute for Physical and Theoretical Chemistry, University of Bonn, 53115 Bonn, Germany. Electronic address: hagelueken@pc.uni-bonn.de.

PMID: 31303480
DOI: 10.1016/j.str.2019.06.007

Abstract

The type-III secretion effector YopO helps pathogenic Yersinia to outmaneuver the human immune system. Injected into host cells, it functions as a Ser/Thr kinase after activation by actin binding. This activation process is thought to involve large conformational changes. We use PELDOR spectroscopy and small-angle X-ray scattering in combination with available crystal structures to study these conformational transitions. Low-resolution hybrid models of the YopO/actin structure in solution were constructed, where the kinase domain of YopO is tilted "backward" compared with the crystal structure, thus shortening the distance between actin and the kinase active site, potentially affecting the substrate specificity of YopO. Furthermore, the GDI domain of the hybrid models resembles a conformation that was previously observed in a crystal structure of the isolated GDI domain. We investigate possible structural reasons for the inactivity of the apo state, analyze its flexibility and discuss the biological implications.

Keywords: DEER; PELDOR; SAXS; Yersinia; YopO; hybrid model; type-III secretion.

Publication types

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

MeSH terms

Actins / metabolism*
Bacterial Proteins / chemistry*
Bacterial Proteins / metabolism*
Catalytic Domain
Models, Molecular
Molecular Docking Simulation
Protein Binding
Protein Conformation
Protein Domains
Scattering, Small Angle
X-Ray Diffraction
Yersinia / chemistry*
Yersinia / metabolism*

Substances

Actins
Bacterial Proteins
YopP protein, Yersinia