Physicochemical bases for protein folding, dynamics, and protein-ligand binding.

Proteins are essential parts of living organisms and participate in virtually every process within cells. As the genomic sequences for increasing number of organisms are completed, research into how proteins can perform such a variety of functions has become much more intensive because the value of the genomic sequences relies on the accuracy of understanding the encoded gene products. Although the static three-dimensional structures of many proteins are known, the functions of proteins are ultimately governed by their dynamic characteristics, including the folding process, conformational fluctuations, molecular motions, and protein-ligand interactions. In this review, the physicochemical principles underlying these dynamic processes are discussed in depth based on the free energy landscape (FEL) theory. Questions of why and how proteins fold into their native conformational states, why proteins are inherently dynamic, and how their dynamic personalities govern protein functions are answered. This paper will contribute to the understanding of structure-function relationship of proteins in the post-genome era of life science research.

A folding "framework structure" of Tetrahymena group I intron.

We have published the dynamic extended folding (DEF) method, which is a RNA secondary structure prediction approach-to simulate the in vivo RNA co-transcriptional folding process. In order to verify the reliability of the method, we selected the X-ray-determined Tetrahymena group I intron as a sample to construct the framework of its folding secondary structure. Our prediction coincides well with the secondary structure predicted by T.R. Cech and the X-ray diffraction crystal structure determined by Lehnert V. Our results show that the DEF framework structure of Tetrahymena group I intron reflects its function sites in a concise and straightforward manner, and the scope of the simulation was expanded.

Cardioviral RNA structure logo analysis: entropy, correlations, and prediction.

In recent years, there has been an increased number of sequenced RNAs leading to the development of new RNA databases. Thus, predicting RNA structure from multiple alignments is an important issue to understand its function. Since RNA secondary structures are often conserved in evolution, developing methods to identify covariate sites in an alignment can be essential for discovering structural elements. Structure Logo is a technique established on the basis of entropy and mutual information measured to analyze RNA sequences from an alignment. We proposed an efficient Structure Logo approach to analyze conservations and correlations in a set of Cardioviral RNA sequences. The entropy and mutual information content were measured to examine the conservations and correlations, respectively. The conserved secondary structure motifs were predicted on the basis of the conservation and correlation analyses. Our predictive motifs were similar to the ones observed in the viral RNA structure database, and the correlations between bases also corresponded to the secondary structure in the database.

Dynamic extended folding: modeling the RNA secondary structures during co-transcriptional folding.

For RNA secondary structure prediction, it is an important issue that how to deal with co-transcriptional folding during the RNA synthesis in the cell. On one hand, co-transcriptional folding, leads to the correct final structure of the whole RNA molecule. On the other hand, it may form the recognition sites for the progress of the transcription. Considering the hurdles in the experimental determination of RNA folding structures, we proposed a so-called "dynamic extended folding simulation" approach. We used two human pre-mRNA samples, the first functional alpha-gene HBZ and the fifth beta-gene HBB, to "display" the co-transcriptional folding images in detail. The modeling process starts from the prediction of a 30-nucleotide (nt) sequence, then in each update 30 nts was extended, say, 1-30, 1-60, 1-90, 1-120,..., 1-1651 nts (for HBB, 1-1606 nts). We selected the RNAstructure program to predict the folding secondary structures of all the segments. We defined "hairpin" as the unit of the secondary structure and analyzed the states of such unit during the sequential dynamic extended folding processes. We found that some hairpins are "conserved", i.e., after its appearance, it always is there in the followed foldings. Some hairpins present partially in the folding segments, and some hairpins appear for only once or twice. This phenomenon vividly depicts the generation and adjusting of the temporal structural units during the co-transcriptional folding process. It is these "hairpins" that support the thermodynamically stable structure at the end of the RNA synthesis. They may also play a role in RNA splicing process and even in the folding structure of the synthesized protein.

[Insight into relationship among mRNA coding region length, folding tendency and codon usage bias in Escherichia coli].

Escherichia coli has been regarded as a model organism in the study of codon usage bias. Most studies in this organism regarding this topic have focused their mind on translation efficiency (translation speed and translation precision) . However, some genes with low codon usage bias and high expression can not be explained by translation efficiency. And some evidence of local RNA secondary structure in coding region of Escherichia coli genes have been given. All of these need explanation from a new point of view. The genomic sequence of Escherichia coli K-12 MG 1655 was obtained from GenBank. Several parameter (Nc, CAI, GC, GC3s) of 4199 genes on codon usage bias were computed. Folding tendency of mRNA coding region was represented by its average energy of 50 nucleotides. And then, relationship among mRNA coding region length, folding tendency and codon usage bias in Escherichia coli were studied. It is argued that though translation efficiency is a primary cause that shapes the codon usage bias of highly expressed genes in Escherichia coli, mRNA coding region length and folding tendency are also unneglectable factor for codon usage bias and weakening of bias. In addition, biological significance of folding tendency in mRNA coding regions was discussed.

Statistical correlation between protein secondary structure and messenger RNA stem-loop structure.

A new integrated sequence-structure database, called IADE (Integrated ASTRAL-DSSP-EMBL), incorporating matching mRNA sequence, amino acid sequence, and protein secondary structural data, is constructed. It includes 648 protein domains. Based on the IADE database, we studied the relation between RNA stem-loop frequencies and protein secondary structure. It was found that the alpha-helices and beta-strands on proteins tend to be preferably "coded" by mRNA stem region, while the coils on proteins tend to be preferably "coded" by mRNA loop region. These tendencies are more obvious if we observe the structural words (SWs). An SW is defined by a four-amino-acid-fragment that shows the pronounced secondary structural (alpha-helix or beta-strand) propensity. It is demonstrated that the deduced correlation between protein and mRNA structure can hardly be explained as the stochastic fluctuation effect.

[The relation between translation speed and protein secondary structure].

Based on the statistical analysis of 119 human and 92 E. coli proteins it was found that for both human and E. coli, the mRNA sequences consisting of tri-codon and tetra-codon with high translation speed preferably code for alpha helices more than for coils. For beta strand, the preference/avoidance oscillates with the translation speed. Moreover, the non-homogeneous usages of tri-codon and tetra-codon with different translation speeds in a given secondary structure have also been found. These results cannot be simply explained by the effect of stochastic fluctuation.

RNAStudio, a full-featured object-oriented program for visualizing RNA secondary structures.

The function of a RNA molecule relies on the underlying physical mechanisms and geometric properties in terms of secondary or tertiary structure, in which one of the most important properties is topological connectivity. RNAStudio, available at http://rnastudio.51.net, brings the convenience of Windows to the representation of RNA folding topology including the sections mentioned below.

Structural analysis of human CCR2b and primate CCR2b by molecular modeling and molecular dynamics simulation.

CCR2b, a chemokine receptor for MCP-1, -2, -3, -4, plays an important role in a variety of diseases involving infection, inflammation, and/or injury, as well as being a coreceptor for HIV-1 infection. Two models of human CCR2b (hCCR2b) were generated by homology modeling and 1 ns restrained molecular dynamics (MD) simulation. In one only C113-C190 forms a disulfide bond (SS model); in another the potential C32-C277 disulfide bond was formed (2SS model). Analysis of the structures and averaged displacements of Calpha atoms of the N-terminal residues shows that the main differences between the SS and 2SS models lie in a region D25YDYGAPCHKFD36; in the extracellular part of the 2SS model the accessible surfaces of N12, F23, Y26, Y28 and F35 are obviously raised and a more stable H-bond net is formed. The potential energy of the 2SS-water assembly finally fluctuated around -43,020 kJ x mol(-1), which is about 302 kJ x mol(-1) lower than that of the SS-water assembly. All these results suggest that the 2SS model is more favorable. The CCR2b genes of 17 primates were sequenced and four CCR2b models for primates Ateles paniscus (A. pan), Hylobates leucogyneus(H. leu), Papio cynocephalus (P. cyn) and Trachypithecus francoist ( T. fra) were generated based on the 2SS model. A comparison of hCCR2b with primate CCR2b also supports the importance of the region D25YDYGAPCHKFD36. Electrostatic potential maps of human and primate CCR2b all display the dipolar characteristics of CCR2b with the negative pole located in the extracellular part and a strong positive pole in the cytoplasmic part. Based on the CCR2b model, we suggest that the main functional residues fall in the D25YDYGAPCHKFD36 region, and the negative electrostatic feature is a non-specific, but necessary, factor for ligands or gp120/CD4 binding.