Amplification and detection of single-molecule conformational fluctuation through a protein interaction network with bimodal distributions.

A protein undergoes conformational dynamics with multiple time scales, which results in fluctuating enzyme activities. Recent studies in single-molecule enzymology have observe this "age-old" dynamic disorder phenomenon directly. However, the single-molecule technique has its limitation. To be able to observe this molecular effect with real biochemical functions in situ, we propose to couple the fluctuations in enzymatic activity to noise propagations in small protein interaction networks such as a zeroth-order ultrasensitive phosphorylation-dephosphorylation cycle. We show that enzyme fluctuations can indeed be amplified by orders of magnitude into fluctuations in the level of substrate phosphorylation, a quantity of wide interest in cellular biology. Enzyme conformational fluctuations sufficiently slower than the catalytic reaction turnover rate result in a bimodal concentration distribution of the phosphorylated substrate. In return, this network-amplified single-enzyme fluctuation can be used as a novel biochemical "reporter" for measuring single-enzyme conformational fluctuation rates.