细胞功能测定

The phosphoinositide cycle is now established as a major pathway by which a variety of hormones, neurotransmitters, and other signaling molecules can generate second messengers and thus affect the intracellular milieu (1). Receptor-mediated activation of membrane-associated ...

It has now been established that agonist stimulation of a large variety of cell-surface receptors promotes hydrolysis of inositol phospholipids through activation of a phosphoinositide-specific phospholipase C (1, 2). Phosphodiesteric cleavage of phosphatidylinositol ...

The ubiquitous intracellular second messengers 3′,5′-cyclic adenosine monophosphate (cAMP) and 3′,5′-cyclic guanosine monophosphate (cGMP) mediate the effects of a large variety of hormones and neurotransmitters. Both of these nucleotides are inactivated by a large group of en ...

Cyclic GMP (cGMP)-dependent protein kinase (G kinase) can be purified from bovine lung with the combination of ion-exchange chromatography, ammonium sulfate precipitation, and cyclic nucleotide affinity chromatography (1). In this chapter, we describe the purification proce ...

A multitude of natural and pharmacological agents, including hormones, neurotransmitters, and phosphodiesterase inhibitors, raise intracellular cAMP levels and, in the vast majority of instances, the physiological effects are the result of activation of cAMP-dependent pr ...

The recent advances in the understanding of the mechanism of action of molecules like nitric oxide and natriuretic peptides have increased the interest in cyclic GMP (cGMP) metabolism. Several techniques have been described to measure cGMP levels that include: 1. A binding assay

Radioligand binding is an extremely versatile technique that can be applied to a wide range of receptors in a variety of preparations, including purified and solubilized receptors, membrane preparations, whole cells, tissue slices, and even whole animals. The basic method is very easy to pe ...

Plasma membranes encompass a complex and varying set of proteins essential to life. In addition, plasma membrane proteins represent the majority of all known drug targets. The characterization of plasma membrane proteomes is, therefore, of eminent importance. A current bottleneck is t ...

A full understanding of leukocyte responses to external stimuli requires knowledge of the full complement of proteins found on their surfaces. Systematic examination of the mammalian cell surfaces at the protein level is hampered by technical difficulties associated with proteo ...

Defining membrane proteomes is fundamental to our understanding of many physiological and pathophysiological processes. Their separation and identification is hence a key issue in basic and biomedical research. Due to their hydrophobic character, few high-resolution techn ...

Platelets (thrombocytes) are the smallest human blood cells and are pivotal in processes of hemostasis and thrombosis. Central to their function, the activation of platelets includes a complex interplay of adhesion and signalling molecules mediated via the plasma and inner membrane. ...

Membranous vesicles are constitutively released by a multitude of cell types. Following fusion of multivesicular bodies with the plasma membrane, endocytic vesicles, 30–90nm in size termed exosomes are released extracellularly. Whilst several groups have reported the presen ...

The modification of cell surface proteins by plasma membrane and soluble proteases is important for physiological and pathological processes. Methods to identify shed and soluble substrates are crucial to further define the substrate repertoire, termed the substrate degradom ...

The discovery of novel target antigens for antibody-based immunotherapy is still a major challenge. Antibody phage display is one of the technologies that is widely applied for the identification of novel cell surface molecules on intact eukaryotic cells and many reports describe the is ...

We identify and describe a set of tools readily available for integral membrane protein prediction. These tools address two problems: finding potential transmembrane proteins in a pool of new sequences, and identifying their transmembrane regions. All methods involve comparing the ...

The social amoeba Dictyostelium discoideum is already known as a model organism for a variety of cellular and molecular studies. Now that the genome sequencing project has been completed and different tools with which to overexpress or knock out genes are available, this species has also moved ...

A DNA microarray consists of an orderly arrangement of DNA, cDNA, or oligonucleotide probes that represent individual genes of an organism. Today, microarrays are the most important tools used to analyze gene transcription on a large scale. Investigations reach from finding key genes in wh ...

In this chapter, we describe how to conveniently demonstrate, assess, and study cell death in Dictyostelium through simple cell culture, clonogenic tests, and photonic (with the help of staining techniques) and electronic microscopy. Cell death can be conveniently generated using mi ...

dictyBase is the model organism database that houses all the sequence and associated data for Dictyostelium discoideum, including literature, researchers, and strains from the Dicty Stock Center. The database makes it possible to connect genes, proteins, and publications and is desi ...

Dictyostelium has a good potential to serve as a model for the study of chromatin function and epigenetic gene regulation. The organism bridges the complexity of higher eukaryotic systems and the simplicity of yeast in that it harbors pathways that are similar to the former and is accessible to ge ...