Premium
Structural characterization of unphosphorylated STAT5a oligomerization equilibrium in solution by small‐angle X‐ray scattering
Author(s) -
Bernadó Pau,
Pérez Yolanda,
Blobel Jascha,
FernándezRecio Juan,
Svergun Dmitri I.,
Pons Miquel
Publication year - 2009
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.83
Subject(s) - small angle x ray scattering , antiparallel (mathematics) , dimer , dissociation (chemistry) , crystallography , chemistry , monomer , biophysics , dissociation constant , intermolecular force , stat protein , protein structure , scattering , stereochemistry , phosphorylation , molecule , biochemistry , biology , stat3 , receptor , physics , organic chemistry , quantum mechanics , magnetic field , optics , polymer
Signal transducer and activator of transcription (STAT) proteins play a crucial role in the activation of gene transcription in response to extracellular stimuli. The regulation and activity of these proteins require a complex rearrangement of the domains. According to the established models, based on crystallographic data, STATs convert from a basal antiparallel inactive dimer into a parallel active one following phosphorylation. The simultaneous analysis of small‐angle X‐ray scattering data measured at different concentrations of unphosphorylated human STAT5a core domain unambiguously identifies the simultaneous presence of a monomer and a dimer. The dimer is the minor species but could be structurally characterized by SAXS in the presence of the monomer using appropriate computational tools and shown to correspond to the antiparallel assembly. The equilibrium is governed by a moderate dissociation constant of K d ∼ 90 μM. Integration of these results with previous knowledge of the N‐terminal domain structure and dissociation constants allows the modeling of the full‐length protein. A complex network of intermolecular interactions of low or medium affinity is suggested. These contacts can be eventually formed or broken to trigger the dramatic modifications in the dimeric arrangement needed for STAT regulation and activity.