Abstract
To photolabel proteins, which are in close proximity to phospholipid head groups on membranes, N-(4-azidosalicyl)phosphatidylethanolamine, ASA-PE, was synthesized. Its special advantage is that it can be radioiodinated in the last step of the synthesis to the [125J]-iodo compound (125J-ASA-PE) with very high specific activity. This is necessary because the4-azido-3[125J]iodo-2-hydroxybenzoyl group is located on the membrane surface and reacts during photolysis as well with proteins as with water. Additionally high specific activity allows to insert low amounts of 125J-ASA-PE into proteoliposomes thus preserving biological activity of their proteins. To study structure and function of the thermophilic ATPsynthase TFo·F1 of the thermophilic bacterium PS 3, it was first reconstituted with soybean phospholipids into liposomes and these were then incubated with 125J-ASA-PE to provide preferred incorporation into the outside of the phospholipid bilayer. After photolysis subunits α of TFo·F1 were labeled selectively. It is, therefore, possible that subunits a may be important in the interaction of TF1 with TFo, to provide internal energy coupling between the proton channel TFo and ATP synthesis in TF1.
Keywords
- High Specific Activity
- Polar Head Group
- Prefer Incorporation
- Saturated Sodium Chloride Solution
- Phospholipid Head Group
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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© 1989 Kluwer Academic Publishers
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Gao, Z., Baeuerlein, E. (1989). 125 J-ASA-PE, A Photoactivable, Radioactive Phospholipid-Analogue, Designed for Photolabeling Proteins in Contact with Phospholipid Head Groups. In: Nielsen, P.E. (eds) Photochemical Probes in Biochemistry. NATO ASI Series, vol 272. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0925-0_5
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DOI: https://doi.org/10.1007/978-94-009-0925-0_5
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-6905-2
Online ISBN: 978-94-009-0925-0
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