生物化学技术

Tyrosine phosphorylation plays a major role in intracellular signal transduction pathways. Phosphorylated tyrosine residues initiate signaling pathways by recruiting proteins containing Src homology-2 (SH2) domains. Herein is described a high-throughput assay to det ...

Recent advances in high-throughput assays for gene expression (genomics), proteins (proteomics), and metabolites (metabolomics) have engendered a parallel need for well-annotated human biological samples. Samples from both diseased and unaffected normal tissues are often ...

Recently, Ciphergen Biosystems (Fremont, CA, USA) has developed a technique called surface-enhanced laser desorption/ionization (SELDI). This technology is based on ProteinChips� with chemically or biochemically modified surfaces for the selective retention and enrich ...

Mass spectrometry (MS)-based proteomic profiling and protein identification has become a powerful tool for the discovery of new disease biomarkers. Among the MS platforms, matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) MS of ...

Laser Capture Microdissection (LCM) is a technique for isolating pure cell populations from a heterogeneous tissue section or cytological preparation through direct visualization of the cells. This technique is applicable to molecular profiling of diseased and disease-free t ...

This chapter describes the rationale behind and means of construction of inexpensive, low to moderate throughput protein arrays. The method of construction is based on injection of analytes into a block of frozen optimum cutting temperature (OCT), the gel media used for frozen sections, and ...

Although the genome provides information about the somatic genetic changes existing in the tissue and underpins pathology, it is the proteins that do the work of the cell and are functionally responsible for almost all disease processes. Moreover, many diseases such as cancer are a manifest ...

As a large number of genome-sequencing projects reached completion, the attention of the scientific community is turning toward understanding the structure-functions of gene translation products—the proteins as well as the complete complement of proteins—the proteome. One go ...

The success of the whole genome sequencing projects brought considerable credence to the belief that high-throughput approaches, rather than traditional hypothesis-driven research, would be essential to structurally and functionally annotate the rapid growth in available ...

Increasing the success in obtaining structures and maximizing the value of the structures determined are the two major goals of target selection in structural proteomics. This chapter presents an efficient and flexible target selection procedure supplemented with a Web-based res ...

Data management has been identified as a crucial issue in all large-scale experimental projects. In this type of project, many different persons manipulate multiple objects in different locations; thus, unless complete and accurate records are maintained, it is extremely difficult to ...

A synopsis of some of the approaches to protein target selection is given, with an emphasis on using web resources to converge on well-ordered, readily crystallizable proteins that are maximally different from known structures. This is illustrated with the genomes of the pathogens causing ...

The Protein Data Bank (PDB) is the repository for the three-dimensional structures of biological macromolecules, determined by experimental methods. The data in the archive are free and easily available via the Internet from any of the worldwide centers managing this global archive. Th ...

The recent advance in our understanding of the relation of protein structure and function cautions that many proteins, or regions of proteins, exist and function without a well-defined three-dimensional structure. These intrinsically disordered/unstructured proteins (IDP ...

Genome sequencing projects have resulted in a rapid increase in the number of known protein sequences. In contrast, only about one-hundredth of these sequences have been characterized using experimental structure determination methods. Computational protein structure mode ...

Domains are considered to be the building blocks of protein structures. A protein can contain a single domain or multiple domains, each one typically associated with a specific function. The combination of domains determines the function of the protein, its subcellular localization and t ...

To fit the requirements of structural genomics programs, new as well as classical methods have been adapted to automation. This chapter describes the automated procedure developed within the Structural Biology and Genomics Platform, Strasbourg for performing recombinant prot ...

For structural studies by x-ray crystallography and nuclear magnetic resonance it is important for the target protein to be available in large quantity and high purity. Escherichia coli expression systems remain the most versatile and convenient means to produce a large quantity of reco ...

Despite the large body of knowledge accumulated on recombinant protein expression, production, primarily of eukaryotic proteins, remains a challenge. The biggest obstacle is in obtaining large amounts of a given protein in a correctly folded form. Several strategies are being used to i ...

The objective of structural proteomics is to determine the structures of all protein folds found in nature and develop a public resource to organize and analyze protein structures and fold families. High throughput (HTP) methods, which can process multiple samples in parallel, saving both ...