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. Author manuscript; available in PMC: 2011 Dec 21.
Published in final edited form as: Mol Microbiol. 2010 Jun 21;77(4):930–942. doi: 10.1111/j.1365-2958.2010.07259.x

Figure 6.

Figure 6

Effect of Hnox1 Fe(II) and Hnox1 Fe(II)-NO on c-di-GMP production by Lpg1057 in vitro. A) 1-D TLC analysis of GTP turnover to c-di-GMP by Lpg1057. Lpg1057 (~ 1 μM) in the presence of Hnox1 (50 μM) in the Fe(II)-NO or Fe(II) unligated states was incubated in sealed anaerobic vials with 500 μM GTP and 10 μCi GTPα32P for 24 hours at 25 °C and then treated with CIP. Arrows indicate the c-di-GMP spot. Spectral analysis of identical reactions without radioactive GTP revealed that less than 10% of the Fe(II) protein was oxidized after 24 hours in the sealed vials, and the Fe(II)-NO complex was stable. B) Relative c-di-GMP production for the Lpg1057 protein alone or in the presence of the Hnox1 Fe(II)-NO or Hnox1 Fe(II). The intensity of the c-di-GMP spots in the phosphorimage were quantified using ImageQuant software (Molecular Dynamics). The error bars represent average values obtained from three independent experiments. *p-values<0.05 for comparison of c-di-GMP production of Lpg1057 with Lpg1057 + Hnox1 Fe(II)NO.