Potential energy surface and molecular dynamics of MbNO: existence of an unsuspected FeON minimum

Ligands such as CO, O(2), or NO are involved in the biological function of myoglobin. Here we investigate the energetics and dynamics of NO interacting with the Fe(II) heme group in native myoglobin using ab initio and molecular dynamics simulations. At the global minimum of the ab initio potential energy surface (PES), the binding energy of 23.4 kcal/mol and the Fe-NO structure compare well with the experimental results. Interestingly, the PES is found to exhibit two minima: There exists a metastable, linear Fe-O-N minimum in addition to the known, bent Fe-N-O global minimum conformation. Moreover, the T-shaped configuration is found to be a saddle point, in contrast to the corresponding minimum for NO interacting with Fe(III). To use the ab initio results for finite temperature molecular dynamics simulations, an analytical function was fitted to represent the Fe-NO interaction. The simulations show that the secondary minimum is dynamically stable up to 250 K and has a lifetime of several hundred picoseconds at 300 K. The difference in the topology of the heme-NO PES from that assumed previously (one deep, single Fe-NO minimum) suggests that it is important to use the full PES for a quantitative understanding of this system. Why the metastable state has not been observed in the many spectroscopic studies of myoglobin interacting with NO is discussed, and possible approaches to finding it are outlined.     

A method for constrained energy minimization of macromolecules

Two algorithms for the local energy minimization of the structure of macromolecules in the presence of constraints are proposed. They are a combination of the method of steepest descents and the method of conjugate gradients with the procedure SHAKE, by which distance constraints can be satisfied. The two algorithms are tested by applying them to a small protein, the bovine pancreatic trypsin inhibitor (BPTI), and compared with the penalty function method for conserving constraints. The efficiency of the proposed methods depends on the level of interdependence of the constraints. For bond-length constraints, the use of SHAKE is superior to the penalty function method. However, when bond-angle constraints are included, SHAKE is more efficient only if the curvature of the penalty function is considerably greater than that of the potential function being minimized. The results indicate that with bond-length constraints the minimization behavior is similar to that without constraints. However, the simultaneous application of bond-length and bond-angle constraints appears to confine the molecule to a very limited part of configuration space, very different from the part covered by an unconstrained minimization. This conclusion calls into question energy minimizations of protein systems in which only the dihedral angles are allowed to vary.     

Vorkommen und Nachweis von Nitriten im Harn

Ohne Zusammenfassung