sHSPs under temperature and pressure: the opposite behaviour of lens alpha-crystallins and yeast HSP26.
Small angle X-ray scattering was used to follow the temperature and pressure induced structural transitions of polydisperse native calf lens alpha-crystallins and recombinant human alphaB-crystallins and of monodisperse yeast HSP26. The alpha-crystallins were known to increase in size with increasing temperature, whereas HSP26 partially dissociates into dimers. SAXS intensity curves demonstrated that the average 40-mer calf alpha-crystallin converted into 80-mer in a narrow temperature range, from 60 to 69 degrees C, whereas the average 30-mer alphaB-crystallin was continuously transformed into 60-mer at lower temperature, from 40 to 60 degrees C. These temperature-induced transitions were irreversible. Similar transitions, yet reversible, could be induced with pressure in the 100 to 300 MPa pressure range. Moreover, temperature and pressure could be combined to lower the transition temperatures. On the other hand, SAXS curves recorded during pressure scans from 0.1 to 200 MPa with monodisperse 24-mer HSP26 revealed dissociation of the 24-mer into dimers. This dissociation was complete and reversible. Whatever the sHSP, a decrease of partial specific volume was found to be associated with the pressure induced quaternary structure transitions, in agreement with the hypothesis that such transitions represent a first step on the protein denaturation pathway.
IMPMC, CNRS-P6-P7 UMR7590, 140 rue de Lourmel, 75015 Paris, France.
Fériel
F
Skouri-Panet
Sophie
S
Quevillon-Cheruel
Magalie
M
Michiel
Annette
A
Tardieu
Stéphanie
S
Finet
D000818:Animals; D002417:Cattle; D006360:Heat-Shock Proteins / Q000737:chemistry*; D007908:Lens, Crystalline / Q000737:chemistry; D011312:Pressure; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D013696:Temperature; D014961:X-Ray Diffraction; D038203:alpha-Crystallin B Chain / Q000737:chemistry*; D038201:alpha-Crystallins / Q000737:chemistry*
C499528:HSP26 protein, S cerevisiae; D006360:Heat-Shock Proteins; D029701:Saccharomyces cerevisiae Proteins; D038203:alpha-Crystallin B Chain; D038201:alpha-Crystallins