Tissue-type plasminogen activator induces alterations in structure and conformation of membrane proteins upon its interaction with human platelets.
In our very recent ESR study we reported that upon rt-PA binding to platelets the H+1/h0 ratios of 16-doxylstearate and 5-doxylstearate spin labels incorporated into the lipid bilayer of platelet membranes were significantly decreased. It corresponded to the increased rigidity of platelet lipid bilayer. In order to further explore this phenomenon we employed a fluorescence-quenching technique which enabled us to estimate the energy transfer efficiency and the apparent interchromophore distance between membrane protein tryptophan and 1-anilino-8-naphthalenesulphonate (ANS) molecules embedded in the membrane lipid bilayer. As t-PA interacts with the platelet membrane this distance decreases, resulting in the relevant increase of energy transfer efficiency. Thus, the data indicate that upon t-PA binding the membrane tryptophan residues are more exposed to the external environment and the quenchable fraction of membrane tryptophan becomes greater. Furthermore, the spectrum of ANS is slightly shifted towards longer wavelengths, which can be accounted for by an increase in the polarity of the environment. It suggests a diminished contact of membrane tryptophan with phospholipid fatty acids. Based on these observations we concluded that the interaction of rt-PA with platelet membranes might induce conformational changes in the membrane proteins, and consequently result in rearrangements of lipid matrix and the alterations in lipid-protein interactions in platelet membranes.
Watala
C
C
Department of Biophysics, Medical University of Lódź, Poland.
Pietrucha
T
T
Dzieiatkowska
E
E
Gwozdinski
K
K
Cierniewski
C S
CS
D001792:Blood Platelets / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D006801:Humans; D008563:Membrane Lipids / Q000097:blood; D008565:Membrane Proteins / Q000097:blood*; D008956:Models, Chemical; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000097:blood; D013050:Spectrometry, Fluorescence; D010959:Tissue Plasminogen Activator / Q000097:blood*
D001798:Blood Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D011994:Recombinant Proteins; D010959:Tissue Plasminogen Activator