Determination of the Hydrophobic Binding Site of Phosphatidylcholine Exchange Protein with Photosensitive Phosphatidylcholine
- 1 September 1979
- journal article
- research article
- Published by Wiley in European Journal of Biochemistry
- Vol. 99 (3), 439-446
- https://doi.org/10.1111/j.1432-1033.1979.tb13274.x
Abstract
1-Acyl-2-{7-(4-azido-2-nitrophenoxy)-[1-14C]heptanoyl}-sn-glycero-3-phosphocholine was synthesized to study the lipid-binding site of the phosphatidylcholine exchange protein from bovine liver. Photosensitive phosphatidylcholine was incorporated into the protein by incubation with vesicles of this phosphatidylcholine derivative. The lipid-protein complex was separated from the vesicles by chromatography on Biogel A-0.5m. Photolysis of the complex by irradiation with light of a high pressure mercury lamp at a wavelength above 340 nm generated the highly reactive nitrene. Sodium dodecyl sulfate gel electrophoresis of the photolysed complex indicated that 30% of the endogenous 14C-labeled phosphatidylcholine was covalently linked to the protein. Peptides were isolated after digestion of the photolysed complex with protease from Staphylococcus aureus and trypsin. The 2-acyl chain of the phosphatidylcholine molecule was linked to the peptide segment -Gly-Ser-Lys-Val-Phe-Met-Tyr-Tyr-. This segment was part of a protease peptide of about 65 residues of which the sequence was determined by Edman degradation for the first 38 residues. This peptide contains a cluster of apolar residues -Val-Phe-Met-Tyr-Tyr-Phe with an extremely high hydrophobicity index and with a predicted .beta.-sheet conformation. This hydrophobic cluster forms part of the binding site.This publication has 35 references indexed in Scilit:
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