Materials

1. SDS-polyacrylamide gel electrophoresis (PAGE) loading buffer: 50 mM Tris-HCl (pH 6.8), 100 mM dithiothreitol, 2% SDS (electrophoresis grade), 0.1% bro-mophenol blue, 10% glycerol.

2. Glass beads: diameter 0.2-0.3 mm (Sigma), acid-washed.

3. Phosphate-buffered saline (PBS) buffer: 136 mM NaCl, 2.6 mM KCl, 10 mM Na2HPO4, 1.76 mM KH2PO4; adjust pH to 7.4 with HCl (9).

4. Isopropylthio-ß-D-galactoside (IPTG): stock solution, 100 mM in water.

6. Lysis buffer: 50 mM NaH2PO4, 300 mM NaCl. Adjust pH to 8.0 using NaOH (10).

7. Wash buffer: 50 mM NaH2PO4, 300 mM NaCl, 20 mM imidazole. Adjust pH to 8.0 using NaOH (10).

8. Elution buffer: 50 mM NaH2PO4, 300 mM NaCl, 250 mM imidazole. Adjust pH to 8.0 using NaOH (10).

9. Interactants (IA) lysis buffer: 50 mM NaH2PO4, 5 mM NaCl. Adjust pH to 7.8 using NaOH.

10. IA wash buffer: 50 mM NaH2PO4, 5 mM NaCl, 20 mM imidazole. Adjust pH to 7.8 using NaOH.

11. IA elution buffer: 50 mM NaH2PO4, 5 mM NaCl, 250 mM imidazole. Adjust pH to 7.8 using NaOH.

12. AffiGel-Hz beads (Bio-Rad).

13. TES-NaOH buffer: 10 mM TES-NaOH, 5 mM NaCl, 5 mM EDTA. Adjust pH to 7.0 using NaOH.

14. IP buffer: 50 mM Tris-HCl, 100 mM KCl, 5 mM MgCl2, 0.2% glycerol, 0.1 mM EDTA.

15. Bovine serum albumin (BSA).

WT CikA- fraction protein

Fig. 1. Immunoanalysis of samples using antisera raised against CikA protein. Lanes 1 and 2: whole cyanobacterial cells (see Subheading 3.1.1.). Lane 3: soluble fraction of wild-type cells (see Subheading 3.2.). Lane 4: His-tagged CikA protein purified from cyanobacterial cells (see Subheading 3.3.1.).

Fig. 1. Immunoanalysis of samples using antisera raised against CikA protein. Lanes 1 and 2: whole cyanobacterial cells (see Subheading 3.1.1.). Lane 3: soluble fraction of wild-type cells (see Subheading 3.2.). Lane 4: His-tagged CikA protein purified from cyanobacterial cells (see Subheading 3.3.1.).

16. SDS-PAGE loading buffer without reducing agent: 50 mM Tris-HCl (pH 6.8), 2% SDS (electrophoresis grade), 10% glycerol.

17. 2-Mercaptoethanol.

18. Bromophenol blue.

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