Extended Data Fig. 1: Sample preparation and characterization of native CrPSI-LHCI-LHCII supercomplex from the pph1;pbcp mutant strain.
From: Structural basis of LhcbM5-mediated state transitions in green algae
a, Sucrose density gradient of solubilized thylakoid membranes. Each band shown is labeled. b, SDS-PAGE analysis of thylakoids (Lane 1) and the purified PSI-LHCI-LHCII supercomplex (Lane 2). The protein composition of each Coomassie band was indicated based on the mass spectrometry and proteomics data analysis. The bands corresponding to PSII subunits are indicated by asterisk. Representative image selected from 3 biological repeats was shown. c, Room-temperature absorption spectra of PSI-LHCI and PSI-LHCI-LHCII samples. The PSI-LHCI-LHCII sample showed higher peaks around 470 and 650 nm (indicated by arrows), demonstrating that the Chl b (from LHCII) content of this fraction is higher than that of PSI-LHCI complex. The spectra were normalized to the maximum in the red region. d, HPLC analysis of pigment content in PSI-LHCI and PSI-LHCI-LHCII samples. Based on the characteristic absorption spectrum of each peak fraction, the six major pigment peaks separated from the sample are identified as loroxanthin/neoxanthin (Lor/Neo), violaxanthin (Vio), lutein (Lut), chlorophyll b (Chl b), chlorophyll a (Chl a) and β-carotene (BCR).
