Filtros Lentos

Fits of the data in each plot were performed using KaleidaGraph v4.01. The data were fit to both single and double-exponentials. Since the data is clearly biphasic in most cases, it was not surprising to that fitting to a double exponential function gave

nucleotide incorporation (Foster, Holmes et al. 2001; Holmes and Erie 2003). Error bars in the rapid quench experiments are reported as standard error, while error bars in the allosteric binding experiments are reported as standard deviation.

In an effort to determine a mechanism to our explain the allosteric loading experiments results, we using KinTekSim (Barshop, Wrenn et al. 1983) to simulate several different kinetic mechanisms. Specifically, we were interested in simulating the extents of [32P]-AMP incorporation as a function of unlabeled ATP concentration. We were able to reduce the field of possible mechanisms down to three kinetically identical mechanisms. There are likely several steps between NTP binding and the actual catalysis event, but we cannot resolve them. The steps presented are the rate limiting steps in each path.

86

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