Figure 6.

Biochemical separation of thylakoid membranes into grana core, grana margins, stroma lamellae, and curvature subfractions. A, A scheme of the classical fractionation of the thylakoid domains obtained with sequential centrifugation at the indicated g values. The curvature region in the grana margin domains is obtained as a loose pellet, distinct from the tight pellet containing the nonappressed stroma lamellae, in the 144,000g step (Puthiyaveetil et al., 2014). B, Representative chl a/b ratio in thylakoid (Thyl) subfractions obtained after solubilization with 0.4% (light gray) or 1% (dark gray) DIG (n = 3). Plants were dark adapted (16 h) before thylakoid isolation. C, Representative immunoblots of the fractions shown in B immunodecorated with α-CURT1B, α-CURT1A, α-PSAB, α-CYT F, and α-D1 antibodies. T, Thylakoids; G, grana core; M, margins; S, stroma lamellae; C, curvature fraction. D, Immunoblot of the supernatant and pellet obtained from dark-acclimated thylakoid membranes with increasing concentrations of DIG and immunodecorated with α-CURT1B, α-CURT1A, α-CYT F (for cyt b6f), α-ATPF (for ATPase), α-D1 (for PSII), α-PSAB (for PSI), and α-LHCB1, α-LHCB2, and α-LHCB3 (for LHCII) antibodies. S, supernatant; P, pellet