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

Biochemistry. 2016 Jan 19;55(2):287-93. doi: 10.1021/acs.biochem.5b01152. Epub 2016 Jan 5.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ferritins / chemistry*
  • Hydrogen-Ion Concentration
  • Scattering, Small Angle*
  • X-Ray Diffraction*

Substances

  • Ferritins