Table 1. Physico-chemical parameters influencing the polymerisation of MamK.

Parameter Influencing Polymerisation	RangeInvestigated	Polymerisation [Scattering Signal]
		Maximum	Minimum
Divalent cations
Calcium ions [CaCl2]	0.125–2 mM	1 mM	2 mM
Magnesium ions [MgCl2]	0.125–2 mM	1 mM	2 mM
Salt
Potassium Chloride [KCl]	0–200 mM	0 mM	200 mM
pH	5.5–9.0	5.5	8.5
Temperature	30–45°C	45°C	30°C

The dependence of polymerisation activity of MamK on divalent metal ion concentration, salt, pH and temperature was monitored by light scattering intensity using a 90° optics scattering angle at a wavelength of 400 nm over a period of 1000 s. All polymerisation reactions were performed using a final concentration of MamK of 10 µM in the assay at 30°C. (a) Divalent metal dependent reactions of MamK were performed in 10 mM imidazole at pH 8.0 and initiated by adding 200 µM ATP containing either calcium or magnesium chloride in the range 0.125–2 mM (b) Salt dependent reactions were performed in 10 mM imidazole at pH 8.0 and initiated by adding 200 µM ATP with varying concentrations of KCl in the range 0–200 mM (100 mM, 50 mM, 25 mM, 12.5 mM, 6.25 mM, 3.125 mM, 1.56 mM) (c) pH dependence of MamK polymerization was performed in 0.1 mM EGTA, 1 mM MgCl2, 20 mM KCl, 200 µM ATP with 10 mM MES at pH 5.5, 10 mM imidazole pH 6.5, 10 mM imidazole pH 7.0, 10 mM imidazole pH 7.0, 10 mM tris pH 7.5, 10 mM tris pH 8.0, 10 mM tris pH 8.5, 10 mM tris pH 9.0 and 10 mM tris pH 9.5 (d) Temperature dependence of MamK polymerization was performed in 10 mM imidazole, 0.1 mM EGTA, 1 mM MgCl2, 20 mM KCl, 200 µM ATP in the range 30–45°C.