Dissociation of cytochrome c from liposomes by histone H1. Comparison with basic peptides.
The basic chromosomal protein histone H1 binds avidly to liposomes containing acidic phospholipids and with characteristics somewhat resembling the lipid association of cytochrome c (cyt c) [Koiv et al. (1995) Biochemistry 34, 8018-8027]. Membrane association of histone H1 strongly attenuates the lipid lateral diffusion in large unilamellar vesicles containing phosphatidylglycerol (PG) as revealed by the decrease in the excimer to monomer ratio Ie/Im of the pyrene fatty acid-containing phospholipid derivative 1-palmitoyl-2-[10-(pyren-1-yl)decanoyl]-sn-glycero-3-phosphogly cer ol (PPDPG) fluorescence. Similarly, an increase in fluorescence anisotropy of the membrane-incorporated probe, diphenylhexatriene (DPH), due to histone H1 indicates that the membrane becomes more rigid. Increasing the mole fraction of PG (XPG) increases in a linear manner the concentration [H1]s required for the maximal decrease in Ie/Im or increase in fluorescence anisotropy, thus allowing to estimate the binding site for H1 to be constituted by approximately 20 PG molecules. Domain formation is also supported by differential scanning calorimetry measurements. Subsequently, we studied the detachment of cyt c from PG-containing liposomes by H1 by measuring its efficiency in decreasing resonance energy transfer between PPDPG and the heme of cyt c. The A-site interaction of 1 microM cyt c with 25 microM PG/PC (XPG = 0.20) liposomes is fully inhibited by low (0.1 microM) histone concentrations. Upon XPG being increased, the concentration [H1]D required for complete detachment of cyt c increases. Irrespective of the [cyt c] present (varying between 0.1 and 10 microM), the C-site-mediated binding of cyt c to neat PG liposomes (XPG = 1.0) is fully prevented at [H1] = 0.6 microM. These measurements indicate that the affinity of histone H1 to liposomes exceeds that of cyt c. The above effects of H1 were subsequently compared with those of different basic membrane-associating peptides. Notably, the effects of HI were remarkably well-reproduced by polylysine (K19). The high affinity of H1 to acidic phospholipids suggests that this feature might also contribute to its physiological function.
Rytömaa
M
M
Department of Medical Chemistry, Institute of Biomedicine, University of Helsinki, Finland.
Kinnunen
P K
PK
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003366:Cosyntropin / Q000737:chemistry / Q000378:metabolism; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism*; D004735:Energy Transfer; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D066298:In Vitro Techniques; D008081:Liposomes / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010715:Phosphatidylglycerols / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D000521:alpha-MSH / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
D003574:Cytochrome c Group; D005456:Fluorescent Dyes; D006657:Histones; D008081:Liposomes; D010446:Peptide Fragments; D010455:Peptides; D010715:Phosphatidylglycerols; D003366:Cosyntropin; D011107:Polylysine; D000521:alpha-MSH