Suplementary Fig. 1aelectrophoretogram of 14% acrylamide gel was conducted for detection of the low molecular weight components of fractions 3-12 of the sucrose gradient shown in Fig. 1. This electrophoresis was stopped when the bromophenol blue arrived at the bottom edge of the gel. The amount of protein in each well was 75 g. In lines 3 and 10 the bands marked with an asterisk (*) were analyzed by LC/ESI/MS/MS and by MALDI-ToF respectively. In line 3 the presence of CpcD2 (gene product of gsr1267) was detected with a 25% coverage and the score of 115 and ApcC (gene product of gsr1244) with a coverage of 78% and the score of 315. In line 10 the RUBISCO small subunit (gene product of glr2158) with a84% coverage and GROES subunit (gene product of glr2896) with a 53% coverage. Panel B shows a 10% acrylamide gel of fractions 3-12 of the sucrose gradient shown in Panel A. This electrophoresis was stopped when the PBPs arrived at the bottom edge of the gel for proper separation of the high molecular weight components. The amount of protein in each well was 75 g except in well 9’ (150 g of protein). The gels were used for Coomassie blue staining and densitometry scanning, to obtain molecular masses and the relative amounts of the proteins. Molecular weight markers were included in each gel, but not shown. The gels were scanned and the apparent molecular masses and relative amounts of the proteins are reported in Supplementary Table 1.Finally the bands marked with an asterisk were subjected to LC/ESI/MS/MS. The data for lines 7 and 9 of Panel B are presented in Tables 1 and 2. The data for line 12 is in the legend of Fig. 1 Panel B. The identified proteins marked with an asterisk in line 8 were: phosphoketolase (gene product of glr0997) with a coverage of 20% and score of 705, multidomain linker (gene product of glr2806) with a coverage of 25% and a score of 744 and nucleotidyltransferase (gene product of gll3149) with a coverage of 22% and score of 707.
Supplementary Fig. 2Sucrose gradient analysis of WCE from G. violaceusPCC7421 after Triton X-100solubilization. FNR activity is expressed as chang in A.U. of DCPIP min-1 ml-1. A shows the FNR activity of fractions obtained from two different sucrose gradients. One gradient was loaded with WCE from G. violaceus after solubilization with Triton X-100 (diamonds in blue ). The second gradient was loaded after the incubation (30 min) of recombinant FNR-3D (1 nmole of FNR) to a equivalent aliquot of a Triton X-100-solubilized WCE from G. violaceus(squares in red ). Also shown (in black triangles ) is the activity that resulted from subtracting the basal activity from the gradient in which the recombinant FNR was added. Panel B show the absorbances at 620 nm (red squares PC) and at 565 nm (blue diamonds PE) and the activity incorporated in the PBPs of G. violaceus(black triangles).
Supplementary Fig. 3.Amino acid sequence of ApcE (LCM130) of G. violaceus. The amino acid sequences of peptide matched by Mascot program are in blue. The score is indicated after the number of the sequence in each row.
ROW 2: 1-78, 2-74, 3-102 ROW 5: 1-94(3) ROW 6: 1-63, 2-55(2)
ROW 7: 65(3) ROW 9: 1-94(2) ROW 10: 1-69 ROW 12: 1-112
ROW 13: 1-83 ROW 14: 1-58, 2-54 ROW 15: 1-88(3) ROW 16: 1-73,
2-59 ROW 17: 1-81(5) ROW 18: 1-50(2) ROW 19: 1-48(2)