Benzophenone-containing fatty acids and their related photosensitive fluorescent new probes: Design, physico-chemical properties and preliminary functional investigations

• Published in: Bioorganic & medicinal chemistry 2011-12, Vol.19 (24), p.7464-7473
• Main Authors: Hilbold, Benoît, Perrault, Marie, Ehret, Christophe, Niu, Song-Lin, Frisch, Benoît, Pécheur, Eve-Isabelle, Bourel-Bonnet, Line
• Format: Article
• Language: English
• Subjects: amino acids; Analytical, structural and metabolic biochemistry; Bacteriorhodopsins - analysis; Benzophenone; Benzophenones - chemical synthesis; Benzophenones - chemistry; Biochemistry, Molecular Biology; Biological and medical sciences; Fatty acid; fatty acids; Fatty Acids - chemical synthesis; Fatty Acids - chemistry; Fluorescence; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Halobacterium salinarum - chemistry; Hydrophobic and Hydrophilic Interactions; hydrophobicity; Life Sciences; Membrane protein; membrane proteins; photoaffinity labeling; Photolabeling; photosensitivity; Proteins; Rhodamines - chemistry; Spectrometry, Fluorescence
• ISSN: 0968-0896; 1464-3391
• DOI: 10.1016/j.bmc.2011.10.043

Hydrophobic photoaffinity labeling is a powerful strategy to identify hydrophobic segments within molecules, in particular membrane proteins. Here we report the design and synthesis of a novel family of fluorescent and photosensitive lipid tools, which have a common amino acid scaffold functionalized by three groups: (i) a first fatty acid chain grafted with a photoactivatable benzophenone moiety (Fatty Acid BenzoPhenone, FABP), (ii) a second fatty acid chain to ensure anchoring into a half-bilayer or hydrophobic environment, and (iii) a fluorescent carboxytetramethylrhodamine headgroup (CTMR) to detect the photolabeled compound. We present data of the synthesis and characterization of three lipid tools whose benzophenone ring is situated at various distances from the central scaffold. We could therefore establish structure/properties relationships dependent upon the depth of insertion of benzophenone into the membrane. Our lipid tools were extensively characterized both physico- and bio-chemically, and we assessed their functionality in vitro using bacterioRhodopsin (bR). We thus provide the scientific community with novel and reliable tools for the identification and study of hydrophobic regions in proteins.