We present an experimental study of the self-assembly of capsid proteins of the cowpea chlorotic mosaic virus (CCMV), in the absence of the viral genome, as a function of pH and ionic strength. In accord with previous measurements, a wide range of polymorphs can be identified by electron microscopy, among them single and multiwalled shells and tubes. The images are analyzed with respect to size and shape of aggregates, and evidence is given that equilibrium has been achieved, allowing a phase diagram to be constructed. Some previously unreported structures are also described. The range and stability of the polymorphs can be understood in terms of electrostatic interactions and the way they affect the spontaneous curvature of protein networks and the relative stabilities of pentamers and hexamers.
The interfacial behavior of asphaltene samples, obtained from two different sources, was studied by means
of Langmuir balance and Brewster angle microscopy (BAM) at the air/water interface. Transferred films from
the air/water interface on mica were studied by atomic force microscopy (AFM). Isotherms obtained from the
Langmuir balance experiments for all samples are qualitatively similar and indicate that their interfacial behavior
may be correlated to the molecular weight of the asphaltene samples. At mesoscopic scales, BAM images of
asphaltene films deposited on the water surface reveal that asphaltene samples do not form a monomolecular
film, as proposed in the literature, but rather a mixture of 2D and 3D domains. This behavior is confirmed
when, on a hydrophilic surface such as mica, the film is transferred from the Langmuir trough and is found
to be not homogeneous or monomolecular. AFM images revealed the formation of an irregular and meandering
layer of 10.2−12.5 nm thickness. Initially, the films exhibit some mesostructures graphically identified as
small and large disklike structures; these disks coalesce or fuse into more complex mesostructures: drumsticklike
and ribbonlike structures. A closer look revealed that these mesostructures are formed by small colloidal-like
particles or small micelles (<100 nm). The spreading capacity of asphaltene on the mica surface seems to be
related to the separation methods and the amount of resins associated with the asphaltenes. The complete
coverage of the surface and the formation of an irregular 3D film occur as a function of aging time or when
the transference pressure of the asphaltene film from water to mica is increased.
The self-assembly of many viral capsids is dominated by protein-protein electrostatic interactions. To have a better understanding of this process, it is important to know how the protein and the capsid surface charges vary as a function of the pH and ionic strength. In this work, using phase analysis light scattering, we measured the electrophoretic mobility (EM) of the cowpea chlorotic mottle virus (CCMV), its capsid protein (CP), and a cleaved CP that lacks its basic terminus, as a function of pH and ionic strength. The EM measurements of the CP are difficult to carry out due to its tendency to self-assemble into capsids; we show how to circumvent this problem by appropriately changing the CP concentration. We found that the isoelectric points (pIs) of the virion and of the CP are insensitive to ionic strength. The onset of multishell structures in the phase diagram of the CCMV CP as a function of ionic strength and pH (and its absence in the brome mosaic virus (BMV) CP phase diagram) can be related to the pI of the capsid. We propose that the transition from multiwall shells to nanotube structures is due to a change in the spontaneous curvature of the CP at its pI. A nonzero limit of the EM at high ionic strength is characteristic of a soft colloid, but a near identity of the EMs of empty capsids and those containing RNA indicates that the EM reflects only the charge distribution in the CP. The Henry equation has been used to provide approximate values of the capsid surface charge as a function of pH and I.
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