生物化学技术

The nuclear inositol lipid cycle is a well known process, and nuclear phosphoinositide-specific phospholipase C β1 (PLCβ1) signalling activity has been extensively studied in the last decades. We now know that nuclear PLCβ1 is a key player in the control of cell cycle progression; in fact it app ...

Nitric oxide synthase (NOS) catalyzes a complex reaction utilizing L-arginine, nicotinamide adenine dinucleotide phosphate (NADPH) and oxygen to synthesize NO, and with citrulline and NADP+ being produced as co-products (reviewed in ref. 1). This chapter describes the measurement ...

Terminal transferase-dependent PCR (TDPCR) can be used after reverse transcription to analyze RNA. This method (RT-TDPCR) is able to provide in vivo information at nucleotide-level resolution, and has been used for study of ribozymes, RNA size, RNA structure, and RNA-protein interacti ...

There is increasing evidence to support the concept that defects in mitochondrial-energy metabolism may underlie the pathology of several disorders such as neurodegenerative diseases. The exact mechanisms involved in the pathogenesis of these disorders are not known, but it is tho ...

Nucleotide analog interference mapping (NAIM) is a quick and effective method to define concurrently, yet singly, the importance of specific functional groups at particular nucleotide residues in relation to the structure and function of an RNA. NAIM can be utilized on virtually any RNA wi ...

The nucleolytic ribozymes are small, autonomously folding RNA species that require metal ions to adopt their active conformations. Three of these ribozymes contain one or more helical junctions, that act as key architectural elements in the structures. A combination of comparative gel ...

In this chapter, we describe a procedure for identification of efficient hammerhead ribozyme (hRz) cleavage sites in target RNAs. An active hRz library, containing randomized recognition sequences flanked by fixed 5′ and 3′ regions, is designed to generate enormous diversity. The libra ...

The methodology for nitric oxide1 (NO) delivery into biological systems has both a biochemical and a pharmacological\toxicological perspective. From the biochemical point of view, the NO delivery system is required to mimic physiological NO production. From the pharmacological ...

Nitric oxide synthases (NOS) are expressed constitutively in vascular endothelial cells (eNOS; 1) and can be induced in vascular smooth muscle following stimulation with cytokines and endotoxin (inducible (i) NOS; for review, see ref. 2). The discovery and characterization of inhibit ...

Nitric oxide synthase (NOS) catalyzes the conversion of L-arginine, molecular oxygen, and nicotinamide adenine dinucleotide phosphate (NADPH) to NO, citrulline, and NADP+ (reviewed in ref. 1). The neuronal (n) and endothelial (e) NOS isozymes are highly regulated by Ca2+ and calmodulin (C ...

In addition to its well-known cofactor roles for aromatic amino-acid hydroxylases, an essential role of tetrahydrobiopterin (BH4) for nitric oxide synthases (NOS) has been recently established (1–3). All three isoforms of NOS contain high-affinity binding sites for BH4. This cofactor ...

Tetrahydrobiopterin (BH4) and related pteridines have received much attention since BH4 was found to be an essential cofactor for nitric oxide synthases (NOS) (1–3). As shown in Fig. 1, BH4 is produced from guanosine 5′-triphosphate (GTP) by three enzymes. GTP cyclohydrolase I converts GTP to d ...

For studies on the biological role of nitric oxide (NO), especially long-term effects of NO on transcriptional or translational regulation of protein expression, it is desired to expose cells to a well-defined NO concentration over a certain period of time. Application of NO gas or NO solutions d ...

Nitration of the ortho position of tyrosine results in the formation of 3-nitrotyrosine. Nitration of tyrosine residues in proteins using tetranitromethane has been used extensively to investigate the role of tyrosine residues in the function of many proteins (1). However, the existe ...

Nitric oxide (NO) has the potential to damage DNA directly, and also to react with O2 to form the nitrosating species N2O3 or to react with O2 − to form the powerful oxidizing agent peroxynitrite (see Chapter 20, this volume). The DNA-damaging activity of NO has been reviewed by Tannenbaum et al. (1).

In 1972, Kerr et al. (1) described a mechanism of cell death that was morphologically distinct from necrosis and they coined the term “apoptosis” , to mean “falling away,” as of leaves from a tree. Although descriptions of cell death consistent with what is now termed apoptosis have been made since 1885 (r ...

Regulation of the inducible nitiric oxide synthase (iNOS) isozyme is distinct from the constitutively expressed endothelial (e) and neuronal (n) NOS isozymes in that it occurs at the level of gene expression and not by Ca+2/calmodulin (CaM) (reviewed in ref. 1). These distinctions have direct ...

Constitutive nitric oxide synthase (NOS) exists as two isoforms: endothelial (e) and neuronal (n) (reviewed in ref. 1). The first reports on the purification of eNOS and nNOS were by Schmidt et al. (2) and Bredt and Snyder (3), respectively; all subsequent reports have been modifications of these proc ...

The authors have developed a fluorescence assay to measure the rate and extent of sterol oxidation in lipid bilayers. Dehydroergosterol (DHE), a fluorescent cholesterol analog, is used as a probe and at the same time as a membrane component. The assay can also be performed on bilayers containing a m ...

Interfacial monolayers used as membrane models have become a practical technique to obtain detailed information about lateral processes taking place in the membrane. These monolayers are particularly useful to study the interactions and parameters governing lateral distri ...