title: Microenvironmental changes in platelet membranes induced by the interaction of fibrinogen-derived peptide ligands with platelet integrins.
abstract: A few studies have confirmed the influence of peptides containing either the RGD or dodecapeptide H-12-V (HHLGGAKQAGDV) sequence on cell membrane structure and function. In order to consider previous findings and to explore microenvironmental changes associated with the interaction of these two fibrinogen-derived peptides with platelet membranes, we employed fluorescence quenching and electron paramagnetic resonance techniques to monitor the possible alterations in platelet membrane dynamics induced by RGDS and H-12-V. The interaction of RGDS with platelet membranes resulted in reduced values of the h+1/ho parameter in both 5-doxylstearic acid and 16-doxylstearic acid spectra indicating a significant rigidification of the membrane lipid bilayer. Otherwise, the fibrinogen-derived peptide that contained the gamma chain C-terminal sequence H-12-V had a fluidizing effect on the platelet membrane lipid bilayer. The labelling of platelet membranes with 1-anilino-8-naphthalenesulphonate (ANS) enabled us to estimate the energy transfer efficiency and the apparent interchromophore distance between membrane protein tryptophan and ANS embedded into the membrane lipid bilayer. As RGDS interacts with platelet membrane this distance decreases, resulting in the relevant increase of energy transfer efficiency. The opposite alterations were recorded upon interaction of platelet membranes with H-12-V. Furthermore, a small shift towards longer wavelengths, which accompanies the spectra of ANS in control platelet membranes, vanishes during the interaction with the peptide H-12-V. This observation can be accounted for by a decrease in the polarity of the ANS environment, and may suggest an enhanced contact of the membrane tryptophan with phospholipid fatty acids. Thus, the data indicate that after the action of H-12-V on platelet membrane receptors, the membrane tryptophan residues become exposed to the external environment and the quenchable fraction of membrane tryptophan becomes smaller. The increase (a) in the relative rotational correlation time (tau c) of 4-(ethoxyfluorophosphinyloxy)-2,2,6,6-tetramethylpiperidine- 1-oxyl (ethoxyfluorophosphinyloxy-TEMPO) and (b) in the hw/hs ratio in the spectra of 4-maleimido-2,2,6,6-tetramethylpiperidine-1-oxyl (maleimido-TEMPO) indicate that under these conditions there is an effective immobilization of some domains located on the hydrated surface of membrane proteins and mobilization of those domains buried inside the membrane protein molecules. The interaction of RGDS with platelet membrane integrins resulted in contrary effects, as compared to H-12-V. In conclusion, our spectroscopic data indicate that these two fibrinogen-derived peptides induce opposite effects in the dynamics of platelet membrane components.
authors.affiliation: Laboratory of Haemostasis and Haemostatic Disorders, Department of Laboratory Diagnostics, Medical University of Lódź, Poland.
authors.forename: C
authors.initials: C
authors.lastname: Watala
authors.forename: K
authors.initials: K
authors.lastname: Gwoźdiński
authors.forename: E
authors.initials: E
authors.lastname: Pluskota
authors.forename: E
authors.initials: E
authors.lastname: Dzieciatkowska
authors.forename: C S
authors.initials: CS
authors.lastname: Cierniewski
mesh_terms: D000595:Amino Acid Sequence; D001792:Blood Platelets / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D005340:Fibrinogen / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D016023:Integrins / Q000378:metabolism*; D008024:Ligands; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence
chemical_list: D016023:Integrins; D008024:Ligands; D008565:Membrane Proteins; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid; D005340:Fibrinogen