化学生物学实验技术

During the past few years, synthetic DNA, used as a primer in DNA polymerization, in site-directed mutagenesis or as a probe in gene selection, has assumed a central role in recombinant DNA technology (1). This was made possible by the development of methods for efficient solid phase chemical synth ...

The initiation of transcription of eukaryotic genes by RNA polymerases is controlled by complex interactions between nonhistone proteins and specific regulatory DNA sequences (promoters and enhancers) (1). In order to characterize and purify such transacting protein factors, a ...

The first successful method for electrophoretic analysis of the full size range of cellular RNA molecules was described by Loening (1), and its introduction allowed for major advances, most particularly in the molecular biology of eukaryotic organisms. The method had, nevertheless, two ...

Computers have become a necessary tool in all laboratories involved in DNA sequence work. There has been a rapid increase in the amount of DNA sequenced, and the most recent issue of the European Molecular Biology Laboratory (EMBL) Nucleotide Sequence Data Library (release 10, 1987) contains a ...

Glycosyltransferases, the enzymes that build oligosaccharides and glycoconjugates, have received much interest in recent years owing to their biological functions and their potential uses in biotechnology. The analysis of the wealth of sequences that are now available in datab ...

β-Galactoside α2,6-sialyltransferase (ST6Gal.I) is the principal sialyltransferase responsible for the biosynthesis of the sialyl α2,6-galactosyl linkage. This enzyme and its cognate glycosidic structure are overexpressed in several malignancies and are related to can ...

The core2 β(1,6)-N-acetylglucosaminyltransferase-I (C2GnT-I) is expressed by leukocytes and is involved in the synthesis of core2 O-glycans that carry sialyl-Lewis x (sLex) oligosaccharides. The core2-based sLex oligosaccharides (C2-O-sLex) have been demonstrated to be phys ...

Glycan molecules covalently linked to proteins or lipids control vital properties of cells, such as signaling, adherence, and migration through the body. The biosynthesis of such glycans depends on the concerted action of many endoplasmic reticulum and Golgi enzymes, a process that is ti ...

Bone cells produce a variety of glycoproteins that contribute to bone health, and function in cell adhesion, stabilizing the extracellular matrix, promoting growth and differentiation, and the induction of apoptosis. Some of these processes appear to be disturbed in arthritis. In this c ...

Methods for rapid and simple analysis of lipopolysaccharide (LPS) from bacterial whole-cell lysates or membrane preparations have contributed to advancing our knowledge of the genetics of the LPS biogenesis. LPS, a major constituent of the outer membranes in Gram-negative bacteria, ...

Gram-negative bacteria have lipopolysaccharides terminating in repeating oligosaccharides which comprise the O-antigen. The glycosyltransferases (GTs) assembling the O-chain utilize lipid-linked acceptor substrates and nucleotide sugar donor substrates. The n ...

Rapid, quantitative methods suited to a large number of samples are required for studies into the determination of disease etiology and in the evaluation of drugs and biological agents. This chapter describes an assay for anionic glycoconjugates (GCs), including glycosaminoglycan ...

Mucins are extremely large and highly O-glycosylated glycoproteins synthesized and secreted by all wet-surfaced epithelia. Within the mucosal secretion they protect the underlying epithelium by forming a selective diffusion barrier against harmful substances and microo ...

Defects in the synthesis of dolichol-linked oligosaccharide (or lipid-linked oligosaccharide ) cause severe, multisystem human diseases called type 1 congenital disorders of glycosylation (CDG type 1). LLOs are also involved in another disease, neuronal ceroid lipofuscinos ...

Despite the extensive structural characterization of glycosphingolipids (GSLs), their functions in cell physiology and pathobiology remain elusive. This is largely owing to the fact that they are difficult to handle, being insoluble in aqueous media, and that no one gene alone determ ...

This chapter describes in detail a practical procedure for the preparation of radiolabeled galactocerebroside and its use in the assay of galactocerebrosidase (GalCase), the enzyme deficient in globoid cell leukodystrophy (Krabbe disease). The reference range for leukocytes a ...

Calnexin and calreticulin are molecular chaperones of the endoplasmic reticulum (ER) whose folding-promoting functions are directed predominantly toward aspargine-linked glycoproteins. This is a consequence of calnexin and calreticulin being lectins with specifici ...

Protein-carbohydrate interactions are believed to be important in many biological processes involving cellular communication. Although many of the experimental approaches for studying protein-carbohydrate interactions are similar to those commonly employed in pro ...

Advances in molecular biology over the last several decades, along with new highly developed methods for protein expression, have enabled investigators to produce and purify large yields of the soluble protein domains of a number of eukaryotic glycosyltransferases and processing g ...

The O-mannosyl glycan is present in a limited number of glycoproteins of brain, nerve, and skeletal muscle. α-Dystroglycan is one of the O-mannosylated proteins and is a central component of the dystrophin-glycoprotein complex that has been shown to be related to the onset of muscular dystrop ...