Comparison of high pressure-induced dissociation of single-stranded DNA-binding protein (SSB) from high pressure-sensitive and high pressure-adapted marine Shewanella species.
The effects of hydrostatic pressure on protein quaternary structure were compared for recombinant single-stranded DNA-binding protein (SSB) derived from piezosensitive, piezotolerant, and obligately piezophilic ("pressure-loving") marine Shewanella strains. The pressure-induced dissociation of the oligomeric SSB proteins was investigated using fluorescence anisotropy. The SSBs all exhibited striking similarity in the pressure-dependent behavior of the fluorescence intensity and emission spectrum as well as in their dissociation constants at atmospheric pressure. The free energies of subunit association into tetramers for all SSBs were between -27 and -30 kcal mol(-1). However, SSB from the piezosensitive Shewanella strain S. hanedai was more sensitive to pressure than that of the SSB proteins from the piezotolerant or piezophilic bacteria. The volume change of association obtained from the pressure dependence of dissociation at a fixed protein concentration (Delta V(p)) for SSB from S. hanedai was 394-402 ml mol(-1). The Delta V(p) values for SSB from the deeper-living Shewanellas were smaller and ranged from 253 to 307 ml mol(-1). Differences between the primary structures of the SSB proteins that could correlate with differences in sensitivity to pressure-induced dissociation were examined.
Chilukuri
Lakshmi N
LN
Center for Marine Biotechnology and Biomedicine, Marine Biology Research Division, Scripps Institution of Oceanography, University of California-San Diego, La Jolla, CA 92093-0202, USA.
Bartlett
Douglas H
DH
Fortes
P A George
PA
D000222:Adaptation, Physiological; D001426:Bacterial Proteins / Q000737:chemistry*; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005454:Fluorescence Polarization; D008970:Molecular Weight; D011312:Pressure; D011489:Protein Denaturation; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D020592:Shewanella / Q000737:chemistry / Q000502:physiology*; D013045:Species Specificity; D013050:Spectrometry, Fluorescence
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins