Surface enhanced resonance Raman scattering as a probe of the spin state of structurally related cytochromes P‐450 from rat liver

FEBS Letters

Smith, W.E.²  Kelly, K.²  Rospendowski, B.N.²  Wolf, C.R.¹ 

[1] Imperial Cancer Research Fund Laboratory of Molecular Pharmacology and Drug Metabolism, Biochemistry Department, George Square, Edinburgh, Scotland;Department of Pure and Applied Chemistry, University of Strathclyde, Cathedral Street, Glasgow G1 1XL Scotland

关键词: Cytochrome P-450;    SERRS;    (Rat liver);   

DOI  :  10.1016/0014-5793(87)80203-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Surface enhanced resonance Raman scattering (SERRS) was observed from structurally related drug-induced rat liver cytochromes P-450 adsorbed on a silver colloid. Careful control of pH and the sequence of addition of components to the sol is required to prevent protein denaturation at the surface due to conversion to P-450's biologically inactive form P-420 or haem loss. A low-spin P-450 (PB_3a), a mixed low-and high-spin P-450 (PB_3b) and a predominantly high-spin P-450 (MC_1a) were investigated. Spectra recorded in the 1300–1700 cm⁻¹ frequency region, containing the oxidation state marker ν₄ at 1375 cm⁻¹ (Fe³⁺) and spin state markers ν₁₀ (1625 cm⁻¹, high-spin; 1633 cm⁻¹, low-spin) and ν₁₉ (1575 cm⁻¹, high-spin; 1585 cm⁻¹, low-spin) were used to differentiate between the spin states of the various forms of cytochrome P-450. As well as the established spin state marker bands, the intensity of a band at 1400 cm⁻¹ appeared to depend on the high-spin content. Thus, with this method SERRS from silver colloids can be used to determine spin states of related cytochromes P-450 in dilute solution (10⁻⁸ M) and may be of value in studies of protein-substrate interactions.

【 授权许可】

Unknown