细胞技术

In this chapter, we describe general methods used to create dynamic computational models of kinase signalling cascades, and tools to support this activity. We focus on the ordinary differential equation models, and show how these fit into a general framework of qualitative and quantitati ...

The convergence and coordinated cross talk of different signalling pathways forms a regulatory network which determines the biological outcome to environmental cues. The MAPK pathways are one of the important routes by which extracellular signals are transduced into intracell ...

Signals transmitted by ERK MAP Kinases regulate the functions of multiple substrates present in the nucleus and the cytoplasm. Once phosphorylated, ERKs dimerize. The functions of these dimers had remained elusive until recently when we demonstrated that ERK dimers are assembled usi ...

ERK cascade scaffolds serve as docking platforms to coordinate the assembly of multiprotein complexes that contribute to the spatial and temporal control of ERK signaling. Given that protein–protein interactions are essential for scaffold function, determining the full reper ...

Signaling through MAPK pathways involves a network of activating kinases and inactivating phosphatases. While single MAPK kinases account for specific activation of the distinct MAPKs, inactivation of MAPKs by phosphatases involves a wider spectrum of enzymes, with phosphata ...

The action of ERK1 and ERK2 activity on the nuclear substrates requires crossing the nuclear envelope and the localization of phospho-ERK into the nucleus. The nucleo-cytoplasmic trafficking of ERK is therefore crucial for the correct functioning of the pathway. Indeed, this step is nece ...

The nuclear–cytoplasmic distribution of ERK2 is regulated in response to various stimuli and changes in cell context. Furthermore, the nuclear flux of ERK2 occurs by several energy- and carrier-dependent and -independent mechanisms. ERK2 has been shown to translocate into and out of the n ...

Mitogen-activated protein (MAP) kinases are a large family of enzymes composed of about four subfamilies, each containing several isoforms and splicing variants. Many MAP kinases are coexpressed in each eukaryotic cell and coactivated in response to various stimuli. It is, therefore, ...

Hydrogen exchange mass spectrometry (HX-MS) is an experimental technique that can be used to �examine solvent accessibility and conformational mobility in biological macromolecules. This chapter summarizes studies using HX-MS to examine the regulation of conformation, pro ...

MAPK cascade components have been the subject of structural analysis, advancing our understanding of how these enzymes are activated and how they interact. A surprising finding has been that unique inactive conformers are adopted by many of these kinases. These inactive conformers are i ...

The present method describes an immunoselection/depletion approach to isolate the native human adipose tissue-derived progenitor cells that are free from endothelial cells and immune cells by the use of magnetic nanobeads and microbeads coupled to antibodies. Moreover, methods ...

The Ras/Raf/MEK/ERK cascade is a highly conserved signal transduction module, whose activation results in a number of different physiological outcomes. Depending on the cell type or the stimulus used, the pathway has been implicated in proliferation, differentiation, apoptosis, a ...

The Ras/mitogen-activated protein kinase (MAPK) pathway is essential in regulation of the cell cycle, cell differentiation, growth, and cell senescence, each of which are critical to normal development. A class of developmental disorders, the “RASopathies,” is caused by germline mut ...

The primary function of the immune system is to protect the organism from invading pathogens. In response to pathogen invasion, multiple signaling pathways are activated in immune cells, leading to diverse immune defense mechanisms. Chief among these pathways is the activation of MAPKs, ...

Vascular Endothelial Growth Factor-A (VEGF-A) is one of the most important regulators of physiological and pathological angiogenesis. Constitutive activation of the ERK pathway and over-expression of VEGF-A are common denominators of tumours of different origins. Understan ...

The mitogen-activated protein kinase (MAPK) family of intracellular signal transducers includes ERK1/2, ERK5, JNK/SAPK, and p38 and has been shown to control survival, proliferation and differentiation of cells composing the central and peripheral nervous system. Some MAPKs pre ...

Maintaining the integrity of the cell cycle is critical for ensuring that cells only undergo DNA replication and proliferation under controlled conditions in response to discrete stimuli. One mechanism by which the fidelity of this process is guaranteed is through the activation of cell ...

Gonadotrophin-releasing hormone (GnRH) is a hypothalamic peptide that acts via Gq/11-coupled 7TM receptors on pituitary gonadotrophs and mediates the central control of reproduction. Recent evidence also indicates that GnRH can affect numerous tissues, but the molecular mech ...

Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Here, we describe methods from our own laboratory and the literature for the isolation and expansion of adipose-d ...

Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Rhesus monkeys are physiologically and phylogenetically similar to humans, and they and their cells are valua ...