Membrane binding induces destabilization of cytochrome c structure.
The effect of membranes binding on the structure and stability of ferricytochrome c was studied by Fourier-transform infrared spectroscopy and differential scanning calorimetry. Association of cytochrome c with phospholipid membranes containing phosphatidylglycerol as a model acidic phospholipid results in only slight, if any, perturbation of the protein secondary structure. However, upon membrane binding, there is a considerable increase in the accessibility of protein backbone amide groups to hydrogen-deuterium exchange, which suggests a lipid-mediated loosening and/or destabilization of the protein tertiary structure. A lipid-induced conformational perturbation of ferricytochrome c is also indicated by a marked decrease in the thermodynamic stability of the membrane-bound protein. Upon binding to membranes containing dimyristoylphosphatidylglycerol (DMPG) or dioleoylphosphatidylglycerol (DOPG) as a single lipid component, the denaturation temperature of ferricytochrome c decreases by approximately 30 degrees C. This is accompanied by a decrease in the calorimetric enthalpy of denaturation, particularly for the DMPG-associated protein. With ferricytochrome c bound to membranes containing a mixture of DMPG (or DOPG) and zwitterionic phosphatidylcholine, the extent of structural perturbation depends on the surface density of the negatively charged lipid head groups, becoming smaller with decreasing proportions of acidic phospholipid in the membrane. The observed destabilization of protein structure mediated by acidic phospholipids (and possibly formation of folding intermediates at the membrane surface) may represent a general property of a larger class of water-soluble proteins for which membrane binding is governed by electrostatic forces.
Muga
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A
Steacie Institute for Molecular Sciences, National Research Council of Canada, Ottawa, Ontario.
Mantsch
H H
HH
Surewicz
W K
WK
D000818:Animals; D002152:Calorimetry, Differential Scanning; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D003574:Cytochrome c Group / Q000378:metabolism*; D005583:Fourier Analysis; D006736:Horses; D008563:Membrane Lipids / Q000378:metabolism*; D010715:Phosphatidylglycerols / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D013055:Spectrophotometry, Infrared; D013696:Temperature; D013816:Thermodynamics
D003574:Cytochrome c Group; D008563:Membrane Lipids; D010715:Phosphatidylglycerols; D010743:Phospholipids; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol; C002773:dimyristoylphosphatidylglycerol