Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1177950 | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics | 2013 | 7 Pages |
We earlier proved the involvement of an autocatalytic step in the oxidation of H2 by HynSL hydrogenase from Thiocapsa roseopersicina, and demonstrated that two enzyme forms interact in this step. Using a modified thin-layer reaction chamber which permits quantitative analysis of the concentration of the reaction product (reduced benzyl viologen) in the reaction volume during the oxidation of H2, we now show that the steady-state concentration of the product displays a strong enzyme concentration dependence. This experimental fact can be explained only if the previously detected autocatalytic step occurs inside the catalytic enzyme-cycle and not in the enzyme activation process. Consequently, both interacting enzyme forms should participate in the catalytic cycle of the enzyme. As far as we are aware, this is the first experimental observation of such a phenomenon resulting in an apparent inhibition of the enzyme. It is additionally concluded that the interaction of the two enzyme forms should result in a conformational change in the enzyme–substrate form. This scheme is very similar to that of prion reactions. Since merely a few molecules are involved at some point of the reaction, this process is entirely stochastic in nature. We have therefore developed a stochastic calculation method, calculations with which lent support to the conclusion drawn from the experiment.
Graphical abstractFigure optionsDownload full-size imageDownload high-quality image (119 K)Download as PowerPoint slideHighlights► Steady-state in the hydrogenase reaction displays enzyme concentration dependence. ► This cannot be explained in the frame of a “normal” enzyme reaction. ► The autocatalytic step should be inside the enzyme cycle. ► Stochastic calculation method succeeded to describe the dependence.