生物化学技术

Heat-shock protein 90 (Hsp90) is a molecular chaperone that assists in the maturation of a limited set of substrate proteins that are collectively referred to as clients. The majority of identified Hsp90 clients are involved in signal transduction, including many steroid hormone recept ...

Heat shock proteins (HSPs) are rapidly induced after stresses, such as heat shock, and accumulate at high concentrations in cells. HSP induction involves a family of heat shock transcription factors that bind the heat shock elements of the HSP genes and mediate transcription in trans. We discu ...

Directed cell migration is fundamental to both physiological and pathophysiological processes such as embryogenesis, wound healing, and cancer metastasis. A complex series of events are required for directional cell migration, which is initiated by a migration-promoting or ch ...

Extracellular heat shock proteins (HSP) play important roles in cell signaling and immunity. Many of these effects are mediated by cell surface receptors expressed on a wide range of cell types. We have investigated the nature of such proteins by cloning candidate receptors into cells (CHO-K ...

Large heat-shock proteins (HSPs), including hsp110 and grp170, are unique immunochaperones capable of carrying and introducing antigens into professional antigen-presenting cells for efficient cross-presentation. Therefore, reconstituted chaperone complexes of l ...

Heat shock proteins (Hsp) are molecular chaperones with the capability to interact with a wide range of other proteins and are thus often found coupled with other heat shock and non-heat shock proteins. This can be an advantage to study specific interactions between a chaperone and other protei ...

Molecular chaperones are a diverse group of highly conserved proteins that transiently interact with partially folded polypeptide chains during normal cellular processes, such as protein translation, translocation, and disassembly of protein complexes (1). Prior to folding ...

Heat-shock protein 90 (HSP90) is an essential molecular chaperone in eukaryotes. It is important for chaperoning proteins that are important determinants of multistep carcinogenesis. HSP90’s ATPase activity is associated with its chaperone function. Co-chaperones as well as po ...

Molecular chaperones promote polypeptide folding in cells by protecting newly made and otherwise misfolded proteins against aggregation or degradation by the ubiquitin proteasome pathway. The roles of Saccharomyces cerevisiae Cdc37 and Ydj1 molecular chaperones are desc ...

The ATPase cycle of Hsp70 chaperones controls their transient association with substrate and, thus, governs their function in protein folding. Nucleotide exchange factors (NEFs) accelerate ADP release from Hsp70 which results in rebinding of ATP and release of the substrate. This cha ...

CHIP, the carboxyl-terminus of Hsp70 interacting protein, is both an E3 ubiquitin ligase and an Hsp70 co-chaperone and is implicated in the degradation of cytosolic quality control and numerous disease substrates. CHIP has been shown to monitor the folding status of the CFTR protein, and we have ...

Human HspB1 (also denoted Hsp27) is a well-known member, together with alphaB-crystallin, of the small heat-shock (or stress) proteins (sHsps) (20–40 kDa). In this chapter, I describe procedures for testing the oligomeric and phosphorylation patterns of HspB1 as well as its interaction with ...

The first step in the affinity purification of antibodies is usually the immobilization of the corresponding antigen to a solid-phase matrix. Generally, this immobilization is done using some chemically reactive gel material such as cyanogen-bromide activated Sepharose. In fact, f ...

Multiple noncovalent forces do play a role in the binding between a protein antigen and an antibody directed against that protein. An antigen is bound by the antigen binding site of an antibody that consists of three hypervariable segments (complementarity determining regions, CDRs) of the ...

Antibodies of the G class can be conveniently purified, even at large scale, by affinity chromatography using immobilized protein A or G. Because specific and cost-effective ligands are not available, scaling up purification of immunoglobin (Ig)M, IgA, and IgE still presents several prob ...

Immunoaffinity chromatography (IAC) has long been regarded as a highly specific method for the purification of biological agents. In this technique, a chromatographic column is used that contains antibodies or antibody-related reagents as the stationary phase. The high selectiv ...

Monoclonal antibodies have many applications in biotechnology such as immuno affinity chromatography, immuno diagnostics, immunotherapy, ding targeting, and biosensors among others. For all these applications, homogeneous antibody preparations are needed. Affinity ...

Recombinant DNA-based manufacturing of biologicals has necessitated the development of processes to recover products from very dilute solutions in high yield and purity. Immunoaffinity chromatography is one such method that takes advantage of the specific and reversible bind ...

In recent years, fluidized-bed adsorption has been used as an alternative method to conventional packed-bed column chromatography for protein purification (1–8). This technology allows the recovery of high-value recombinant proteins and other biomolecules straight from unc ...

Since the inception of affinity chromatography 30 years ago (1), it has developed into a powerful tool mainly for the purification of proteins. It is based on the reversible formation of a tight binding complex between a ligand, immobilized on an insoluble matrix and a substance, the ligate, to be isol ...