Conformational changes preceding decapsidation of bromegrass mosaic virus under hydrostatic pressure: A small-angle neutron scattering study

Autor(en): Leimkuhler, M
Goldbeck, A
Lechner, MD
Witz, J
Stichwörter: Biochemistry & Molecular Biology; bromegrass mosaic virus; CHLOROTIC MOTTLE VIRUS; COAT PROTEIN; DENATURATION; DISSOCIATION; GLYCOPROTEIN; IONIZATION; PH; POLIOVIRUS; pressure effects; RESOLUTION; small-angle neutron scattering; STABILITY; swelling
Erscheinungsdatum: 2000
Volumen: 296
Ausgabe: 5
Startseite: 1295
Seitenende: 1305
The stability of bromegrass mosaic virus (BMV) and empty shells reassembled in vitro from purified BMV coat protein was investigated under hydrostatic pressure, using solution small-angle neutron scattering. This technique allowed us to monitor directly the dissociation of the particles, and to detect conformational changes preceding dissociation. Significant dissociation rates were observed only if virions swelled upon increase of pressure, and pressure effects became irreversible at very high-pressure in such conditions. At pH 5.0, in buffers containing 0.5 M NaCl and 5 mM MgCl2, BMV remained compact (radius 12.9 nm), dissociation was limited to approximate to 10 % at 200 MPa, and pressure effects were totally reversible. At pH 5.9, BMV particles were slightly swollen under normal pressure and swelling increased with pressure. The dissociation was reversible to 90% for pressures up to 160 MPa, where its rate-reached 28%, but became totally irreversible at 200 MPa. Pressure-induced swelling and dissociation increased further at pH 7.3, but were essentially irreversible. The presence of (H2O)-H-2 in the buffer strongly stabilized BMV against pressure effects at pH 5.9, but not at pH 7.3. Furthermore, the reversible changes of the scattered intensity observed at pH 5.0 and 5.9 provide evidence that pressure could induce the release of coat protein subunits, or small aggregates of these subunits from the virions, and that the dissociated components reassociated again upon return to low pressure. Empty shells were stable at FH 5.0, at pressures up to 260 MPa. They became ill-shaped at high-pressure, however, and precipitated slowly after return to normal conditions, providing the first example of a pressure-induced conformational drift in an assembled system. (C) 2000 Academic Press.
ISSN: 00222836
DOI: 10.1006/jmbi.2000.3538

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