Interaction of mammalian Hsp22 with lipid membranes.
Hsp22/HspB8 is a member of the small heat-shock protein family, whose function is not yet completely understood. Our immunolocalization studies in a human neuroblastoma cell line, SK-N-SH, using confocal microscopy show that a significant fraction of Hsp22 is localized to the plasma membrane. We therefore investigated its interactions with lipid vesicles in vitro. Intrinsic tryptophan fluorescence is quenched in the presence of lipid vesicles derived from either bovine brain lipid extract or purified lipids. Time-resolved fluorescence studies show a decrease in the lifetimes of the tryptophan residues. Both of these results indicate burial of some tryptophan residues of Hsp22 upon interaction with lipid vesicles. Membrane interactions also lead to increase in fluorescence polarization of Hsp22. Gel-filtration chromatography shows that Hsp22 binds stably with lipid vesicles; the extent of binding depends on the nature of the lipid. Hsp22 binds more strongly to vesicles made of lipids containing a phosphatidic acid, phosphatidylinositol or phosphatidylserine headgroup (known to be present in the inner leaflet of plasma membrane) compared with lipid vesicles made of a phosphatidylcholine head-group alone. Far-UV CD spectra reveal conformational changes upon binding to the lipid vesicles or in membrane-mimetic solvent, trifluoroethanol. Thus our fluorescence, CD and gel-filtration studies show that Hsp22 interacts with membrane and this interaction leads to stable binding and conformational changes. The present study therefore clearly demonstrates that Hsp22 exhibits potential membrane interaction that may play an important role in its cellular functions.
Chowdary
Tirumala Kumar
TK
Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India.
Raman
Bakthisaran
B
Ramakrishna
Tangirala
T
Rao
Ch Mohan
ChM
D000818:Animals; D002850:Chromatography, Gel; D006360:Heat-Shock Proteins / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism*; D011487:Protein Conformation / Q000187:drug effects; D051381:Rats; D013050:Spectrometry, Fluorescence
D006360:Heat-Shock Proteins; C490753:HspB8 protein, rat; D008081:Liposomes; D008563:Membrane Lipids