The detergent and salt effect on the light-harvesting chlorophyll a/b complex from green plants.
The light harvesting accessory pigment-protein complex (LHC) with a chlorophyll (Chl) a/b ratio of 1.2 was isolated by treating pea chloroplasts with Triton X-100. The LHC was used to investigate the action of ionic (sodium dodecyl sulfate) and non-ionic (Triton X-100) detergents. By optical methods (absorption and fluorescence spectra, measurements of fluorescence yield, phi, and lifetime, tau) two successive stages of the process were demonstrated, namely (1) interaction between detergent monomers and proteins and (2) solubilization of pigments into detergent micelles, which is facilitated by the presence of salts. The concentration ranges, characteristic of these stages, differ by 1.5-2 orders of magnitude for SDS, but slightly overlap for Triton X-100. At the second stage, certain changes occur in LHC absorption and fluorescence spectra. Several stable states of the LHC were established: (1) an aggregated state formed the presence of 10 mM MgSO4 with tau approximately 0.6 ns; (2) the dialyzed LHC with tau approximately 0.9 ns; (3) the states of the LHC in detergent solution with tau approximately 2.3, 2.9, 3.4 ns; (4) a 30 kilodalton monomer obtained by SDS-polyacrylamide gel electrophoresis with tau approximately 4.1 ns. The fluorescence parameters of the LHC states were compared with those of Chl a in detergent micelles (for the micelles tau = 5.6-6.0 ns). The tau/phi ratio (the criterion for emission heterogeneity) for the LHC in the absence of a detergent was shown to be higher at least by a factor of 3.5 than that for Chl a in the presence of a detergent. Successive additions of the detergent to the LHC cause gradual decrease in the tau/phi ratio, and for the LHC monomer it reaches practically the same value as for Chl a in detergent micelles. The results are discussed on the basis of the data obtained previously, It is suggested that in vivo LHCs do not form such aggregates as in water solution without a detergent.
D000042:Absorption; D002734:Chlorophyll / Q000378:metabolism*; D008278:Magnesium Sulfate / Q000494:pharmacology; D017830:Octoxynol; D010944:Plants / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D002734:Chlorophyll; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D008278:Magnesium Sulfate; D017830:Octoxynol