The open-bundle structure of cytochrome c’ as an unfolding intermediate was determined by small-angle neutron scattering experiment (SANS). The four-α-helix bundle structure of Cyt c’ at neutral pH was transited to an open-bundle structure (at pD ~13), which is a joint-clubs consisting of four clubs (α-helices) connected by short loops. The compactly folded structure of Cyt c’ (radius of gyration, Rg = 18 Å for the Cyt c’ dimer) at neutral or mildly alkaline pD transitioned to a remarkably larger “open-bundle” structure at pD ~13 (Rg = 25 Å for the Cyt c’ monomer). Cyt c’ adopts an unstructured random coil structure at pD = 1.7 (Rg = 25 Å for the Cyt c’ monomer). Numerical partial scattering function analysis (joint-clubs) and ab initio modelling gave structures similar to the “open-bundle”, which retains the α-helices but loses the bundle structure.

In this manuscript we are investigating the contribution of dynamic membrane properties of phospholipid membranes to coherent scattering signals under grazing incidence. Spectroscopic measurements under grazing incidence can provide useful insight into the properties of biological membranes, however are often impeded by weak signals. By using grazing-incidence small-angle neutron scattering (GISANS) to identify a dynamic scattering contribution we are able to independently corroborate the existence of a previously found dynamic mode found by grazing-incidence neutron spin echo spectroscopy (GINSES). Additionally, by increasing the speed of measurement compared to GINSES from several days to hours we were able to explore the temperature behavior of this mode in phospholipid membranes. These dynamic modes of the membranes show a wavelength of around 700 Å in-plane of the membrane and are most pronounced around 37°C and strongly decrease at lower temperatures below 25°C before vanishing at 20°C. We therefore speculate that they may be linked to biologically relevant functions of the membranes themselves.