Figure 1.
Pattern of thylakoid membrane complexes separated from rice by 1D BN-PAGE.
A: The lanes 1, 2, 3, 4, and 5 were loaded with preparations of 50 µg chlorophyll(∼500 µg protein) dissolved in 50 µl of 0.5, 1.0, 2.0, 3.0, and 4.0% DDM lyses buffers. B: Each slot of lanes 1, 2, and 3 was loaded with the supernatants of 50 µg chlorophyll(∼500 µg protein) dispersed corresponding to 50 µl 4% NP-40, TritonX-100, and DDM lyses buffers. C: Lane I was loaded with high MW markers dissolved in 50 µL 4.0% DDM lyses buffers and electrophoresed in the same condition with Lane II. Lane II was loaded with the supernatant of 50 µg chlorophyll dispersed in 50 µL 4.0% DDM lyses buffers and scanned directly after electrophoresis, and lane III was the same as lane II and scanned after stained by Coomassie Blue, nine membrane complexes were labelled on the left of Coomassie-stained gel.
Table 1.
Apparent MW of thylakoid membrane complexes from rice (Oryza sativa L.).
Figure 2.
The operation of a novel “Double-Strips 2D BN/SDS-PAGE” transfer technique from the first dimension to the second.
A: One strip of 1D BN-gel lane was placed in the glass plate after denaturing. B: The other identical BN-gel strip was covered to the previous according to the aligned protein complexes, and then the glass plate was overlapped on the gels. C: The cassette was placed in skew form avoiding air bubble produced, the agarose was poured in the space between the BN-gel and stacking gel (just the half of all volume, as arrow indicated) after acrylamide was polymerized. D: The remaining space was subsequently filled with agarose until the previous solution has been concreted.
Figure 3.
Resolution comparison between “single-strip BN/SDS-PAGE” and "double-strips BN/SDS-PAGE” with membrane proteins.
A: Comparison of 2D gels pattern between “Single-strip BN/SDS-PAGE” and “Double-strips BN/SDS-PAGE” in rice. (D1): The pattern of “Double-Strips BN/SDS-PAGE” with two 1D BN-Gel lane strips each loaded with 50 µg chlorophyll dissolved at DDM/protein ratio of 4/1. (S1): The pattern of “Single-Strip BN/SDS-PAGE” with one 1D BN-Gel strip loaded with 100 µg chlorophyll dissolved at DDM/protein ratio of 4/1. (S2): The pattern of “Single-Strip BN/SDS-PAGE” with one 1D BN-Gel strip loaded with 50 µg chlorophyll dissolved at DDM/protein ratio of 4/1. (Parts of regions α, β and γ in the panels of 2D SDS-gels represent the obvious difference between these methods). B: The pattern of the thylakoid membrane proteins from Spinacia oleracea and Cucurita spp. separated by “Double-Strips BN/SDS-PAGE”. The amounts of chlorophyll and detergent/protein ratios were labelled in the panels.
Figure 4.
The constituent subunit pattern of thylakoid membrane complexes in rice (Oryza sativa L.) resolved using “Double-Strips BN/SDS-PAGE” (Each protein spot was marked with capital letter plus Roman numeral, which was corresponded to the spot mark in Table 2.)
Table 2.
Identification of thylakoid membrane proteins in rice (Oryza sativa L.) by MALDI-TOF.
Figure 5.
The modelled structure of the PsaC, PsaD and PsaE from rice (Oryza sativa L.).
A: Two important salt bridge interactions were labelled on the model. B: The conformation change of the salt bridge during the 15 ns simulation.
Figure 6.
Characterization of stromal ridge complex in explicit solvent MD.
A: The RMSD of the PsaC, PsaD and PsaE for the 15 ns MD run. B: Computational alanine scanning mutagenesis results for six single mutations from Oryza sativa, Synechococcus elongates and Pisum sativum. C: The closest atom distance in the residue pairs R19-E168, R75-V89, R75-E90, R75-E91. D: Interaction energy in the residue pairs R19-E168, R75-V89, R75-E90, R75-E91.