The preparation of all-trans uniformly (13)C-labeled retinal via a modular total organic synthetic strategy. emerging central contribution of organic synthesis toward the structure and function study with atomic resolution in protein research

Uniformly [(13)C(20)]-labeled all-trans-retinal (1) has been prepared via a convergent modular total organic strategy with high isotope incorporation (>99%) and without isotope dilution starting from commercially available 99% enriched (13)C-labeled starting materials. For this purpose we have developed a strategy that is based on four different modules: [1,2,3,4,(3-CH(3))-(13)C(5)]-4-(diethylphosphono)-3-methyl-2-butenenitrile (3), [1,2,3,4-(13)C(4)]-ethyl acetoacetate (7), [U-(13)C(5)]-4-bromo-2-methyl-2-butene (13), and [U-(13)C(10)]-2,6,6-trimethylcyclohex-2-ene-1-ylcarbonitrile (16). This scheme permits the synthesis of the full cassette of all isotopomers with (13)C-labels at any position or combination of positions by using different (13)C-labeled starting materials. In addition, modifications of the synthesized modules will give access to a broad range of chemically modified (13)C-labeled retinoids and carotenoids. This modular strategy enables the synthesis of multifold and uniformly stable isotopically labeled (bio)macromolecules that can be used for studying proteins with atomic resolution, providing detailed functional information of the studied biological system.