De novo protein structure modeling from cryoem data through a dynamic programming algorithm in the secondary structure topology graph.

摘要

Proteins are the molecules carry out the vital functions and make more than the half of dry weight in every cell. Protein in nature folds into a unique and energetically favorable 3-Dimensional (3-D) structure which is critical and unique to its biological function. In contrast to other methods for protein structure determination, Electron Cryo-rricroscopy (CryoEM) is able to produce volumetric maps of proteins that are poorly soluble, large and hard to crystallize. Furthermore, it studies the proteins in their native environment. Unfortunately, the volumetric maps generated by current advances in CryoEM technique produces protein maps at medium resolution about (~5 to 10Å) in which it is hard to determine the atomic-structure of the protein. However, the resolution of the volumetric maps is improving steadily, and recent works could obtain atomic models at higher resolutions (~3Å).;De novo protein modeling is the process of building the structure of the protein using its CryoEM volumetric map. Thereupon, the volumetric maps at medium resolution generated by CryoEM technique proposed a new challenge. At the medium resolution, the location and orientation of secondary structure elements (SSE) can be visually and computationally identified. However, the order and direction (called protein topology) of the SSEs detected from the CryoEM volumetric map are not visible. In order to determine the protein structure, the topology of the SSEs has to be figured out and then the backbone can be built. Consequently, the topology problem has become a bottle neck for protein modeling using CryoEM.;In this dissertation, we focus to establish an effective computational framework to derive the atomic structure of a protein from the medium resolution CryoEM volumetric maps. This framework includes a topology graph component to rank effectively the topologies of the SSEs and a model building component. In order to generate the small subset of candidate topologies, the problem is translated into a layered graph representation. We developed a dynamic programming algorithm (TopoDP) for the new representation to overcome the problem of large search space. Our approach shows the improved accuracy, speed and memory use when compared with existing methods. However, the generating of such set was infeasible using a brute force method. Therefore, the topology graph component effectively reduces the topological space using the geometrical features of the secondary structures through a constrained K-shortest paths method in our layered graph. The model building component involves the bending of a helix and the loop construction using skeleton of the volumetric map. The forward-backward CCD is applied to bend the helices and model the loops.