- Shape and lipid binding site of the nonspecific lipid-transfer protein (sterol carrier protein 2): a steady-state and time-resolved fluorescense study
- Pages (from-to)
- Number of pages
- Document type
- Faculty of Science (FNWI)
- Swammerdam Institute for Life Sciences (SILS)
The nonspecific lipid-transfer protein (nsL-TP) from bovine liver was studied with time-resolved and steady-state fluorescence techniques. From the decay of the intrinsic tryptophanyl fluorescence, it wasestimated that the rotational correlation time of nsL-TP is 15 ns. This parameter increased only slightly upon addition of an excess of negatively charged vesicles, indicating that the basic nsL-TP is not immobilized at the membrane surface under these conditions. Binding studies using fluorescent lipid analogues revealed that nsL-TP is able to extract sn-2-(pyrenehexanoyl)phosphatidylcholine and l-palmitoyl-2-[ 34di-phenylhexatrienyl)propionyl]-sn-3-phosphocholine (DPHp-PC) from a quenched donor vesicle. The fluorescence increase resulting from this binding was poorly quenched by either acrylamide or iodide. This indicates that nsL-TP shields the bound PC molecules from the aqueous environment. Timeresolved analysis of DPH fluorescence originating from DPHp-PC bound to nsL-TP yielded a rotational correlation time of 7.4 ns. This correlation time strongly suggests that the DPH moiety of the bound molecule is immobilizedand that the nsL-TP/DPHp-PC complex is not attached to the donor vesicle. In view of the longer rotational correlation time obtained for the intrinsic tryptophanyl fluorescence, we conclude that nsL-TP is highly asymmetric. The data are consistent with a model in which the shape of nsL-TP is ellipsoidal with an axis ratio of 2.8. The implications for the mode of action of nsL-TP are discussed.
- go to publisher's site
If you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library, or send a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible.