Ferritin Assembly Revisited: A Time-Resolved Small-Angle X-ray Scattering Study.

The assembly reaction of Escherichia coli ferritin A (EcFtnA) was studied using time-resolved small-angle X-ray scattering (TR-SAXS). EcFtnA forms a cagelike structure that consists of 24 identical subunits and dissociates into dimers at acidic pH. The dimer maintains nativelike secondary and tertiary structures and is able to reassemble into a 24-mer when the pH is increased. The reassembly reaction was induced by pH jump, and reassembly was followed by TR-SAXS. Time-dependent changes in the forward scattering intensity and in the gyration radius suggested the existence of a significant population of intermediate oligomers during the assembly reaction. The initial reaction was a mixture of second- and third-order reactions (formation of tetramers and hexamers) from the protein concentration dependence of the initial velocity. The time-dependent change in the SAXS profile was roughly explained by a simple model in which only tetramers, hexamers, and dodecamers were considered as intermediates.

[1]  Xiaohong Liu,et al.  A trimer of dimers is the basic building block for human immunodeficiency virus-1 capsid assembly. , 2012, Biochemistry.

[2]  E. Chiancone,et al.  On the mechanism of horse spleen apoferritin assembly: a sedimentation velocity and circular dichroism study. , 1987, Biochemistry.

[3]  Marileen Dogterom,et al.  Microtubule organization in vitro. , 2013, Current opinion in cell biology.

[4]  Arnau Casañas,et al.  Vault particles: a new generation of delivery nanodevices. , 2012, Current opinion in biotechnology.

[5]  P. Harrison,et al.  Self-assembly of apoferritin from horse spleen after reversible chemical modification with 2,3-dimethylmaleic anhydride. , 1988, Biochemistry.

[6]  Todd O Yeates,et al.  The protein shells of bacterial microcompartment organelles. , 2011, Current opinion in structural biology.

[7]  S. Stahl,et al.  A theoretical model successfully identifies features of hepatitis B virus capsid assembly. , 1999, Biochemistry.

[8]  Yu Zhang,et al.  Self-Assembly in the Ferritin Nano-Cage Protein Superfamily , 2011, International journal of molecular sciences.

[9]  B. Schmitt,et al.  Performance of single-photon-counting PILATUS detector modules , 2009, Journal of synchrotron radiation.

[10]  G. Tresset,et al.  Norovirus capsid proteins self-assemble through biphasic kinetics via long-lived stave-like intermediates. , 2013, Journal of the American Chemical Society.

[11]  R. Jaenicke,et al.  Mechanism of the self-assembly of apoferritin from horse spleen , 1987, European Biophysics Journal.

[12]  A. P. Hammersley,et al.  Two-dimensional detector software: From real detector to idealised image or two-theta scan , 1996 .

[13]  Mauricio G Mateu,et al.  Assembly, stability and dynamics of virus capsids. , 2013, Archives of biochemistry and biophysics.

[14]  Daisuke Sato,et al.  A Physicochemical and Mutational Analysis of Intersubunit Interactions of Escherichia coli Ferritin A. , 2015, Biochemistry.

[15]  T. C. Huang,et al.  X‐ray powder diffraction analysis of silver behenate, a possible low‐angle diffraction standard , 1993 .

[16]  Roman Tuma,et al.  Detection of intermediates and kinetic control during assembly of bacteriophage P22 procapsid. , 2008, Journal of molecular biology.

[17]  K. Hughes,et al.  Coordinating assembly of a bacterial macromolecular machine , 2008, Nature Reviews Microbiology.

[18]  G. Fagherazzi.  Small angle X‐ray scattering edited by O. Glatter and O. Kratky , 1983 .

[19]  T. Uruga,et al.  Small‐angle X‐ray scattering station at the SPring‐8 RIKEN beamline , 2000 .

[20]  D. Svergun,et al.  CRYSOL : a program to evaluate X-ray solution scattering of biological macromolecules from atomic coordinates , 1995 .

[21]  T. J. Stillman,et al.  The high-resolution X-ray crystallographic structure of the ferritin (EcFtnA) of Escherichia coli; comparison with human H ferritin (HuHF) and the structures of the Fe(3+) and Zn(2+) derivatives. , 2001, Journal of molecular biology.

[22]  Uri Raviv,et al.  RNA encapsidation by SV40-derived nanoparticles follows a rapid two-state mechanism. , 2012, Journal of the American Chemical Society.