更多资讯

Retinoic acid receptors (RARs) have been detected in nuclear extracis from various cell types on the basis of their ability to bind radiolabeled retinoic acid (1–3). Prior to the isolation of RAR cDNAs in 1987 (4, 5), biochemical evidence for the existence of nuclear retinoic-acid binding protei ...

In situ hybridization (ISH) is a technique that enables one to localize an endogenous mRNA in histologtcal and cytological samples to the resolution of nearly single cells. For this reason, ISH is widely used in the fields of genetics, developmental biology, and neurobiology, as well as for dtagn ...

Retinoids, a group of structural and functional analogs of vitamin A and clinically important agents for chemoprevention, can modulate epithelial differentiation and suppress carcinogenesis in various tissues, including skin, bladder, oral cavity, lung, and mammary gland in ex ...

Ligand-binding assays have been used extensively in the past to detect and characterize intracellular receptors. In fact, the mitial discoveries of the first known members of this family of proteins were achieved by the demonstration of binding of radioactively labeled hormonal liga ...

Techniques for the preparation of antibodies and their applications are widely established in the literature. Antibodies have proven to be useful reagents in characterizing the molecular activities of the retinoid receptors (1). The use of a retinoid “X” receptor α (RXRα) antibody on Wes ...

Antisense methodologies have been used extensively to inhibit the expression of specific genes with a view to elucidating their role in particular cellular processes (1). The technique is based on the ability of mRNA to bind, in a sequence-specific fashion, to a complimentary oligonucleo ...

Information regarding rates of dissociation of ligands from binding proteins is helpful for obtaining insights into the forces that stabilize protein-ligand interactions, as well as for understanding how the ligands distribute between the different cellular compartments, w ...

Partial proteolysis of an undenatured protein is a wtdely used, powerful technique to probe protein conformation in the native state. The basts for thts technique is that the more exposed an amino-acid residue is to the solvent, the easier it 1s for a protease to cleave a peptide bond at that site (1, 2). Ther ...

To study the interaction of retinol-binding protein (RBP) with its plasma carrier, transthyretin (TTR), spectrofluorimetry, and circular dichroism have previously been used. Both these techniques require milligram quantities of the proteins and this sets limitations on the use of ...

Vitamin A is transported in the plasma as retinol bound to a carrier protein, called retinol-binding protein (RBP), which itself forms a complex with the thyroxine-binding protein, known as transthyretin (TTR). This complex exists in equilibrium with free holo-RBP, which can then interact w ...

Fluorescence spectroscopy has long been used to characterize the equilibrium binding of retinoids to proteins (1). Changes in steady-state fluorescence are monitored as the protein is titrated with aliquots of retinoid. The resultant binding curve can be analyzed yielding informa ...

The intracellular carriers for all-trans retinol are believed to be the homologous cellular retinol-binding protein (CRBP) and cellular retinol-binding protein II (CRBP II) (ref. 1, and references therein). CRBP is present in a wide variety of tissues, including liver, kidney, and testis. ...

Nuclear-retinoid receptors provide a mechanism of retinoid action, but most likely where expressed, the cytosolic cellular retinoic acid-binding proteins (CRABPs) also affect the ability of retinoids to initiate biological signals (1, 2). holo-CRABP I sequesters retinoic acid ( ...

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) result in defective ion transport, leading to thick mucus, impaired mucociliary clearance and decreased bacterial killing in the lung (1,2). Despite recent progress in associating cystic fibrosis (CF) ...

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene are responsible for cystic fibrosis (CF) (1) and the primary defect in CF is the loss of cAMP-regulated anion conductance in the apical plasma membrane of epithelial cells in affected tissues (2,3). Primary c ...

On April 17, 1993, Crystal and co-workers initiated the first human gene therapy trial for cystic fibrosis (CF). In that study, a replication-deficient recombinant adenovirus encoding cystic fibrosis transmembrane conductance regulator (CFTR) cDNA was administered to the nasal and ...

Airway surface liquid (ASL) is generally described as a two-layer system formed by a “sol” (or periciliary) and a “gel” layer (overlying the cilia). The ASL, whose total thickness is estimated to be between 10 and 40 �m, contributes to mechanical and antimicrobial defense systems in host airways. In cy ...

Following the identification and subsequent cloning of the gene that is defective in patients with cystic fibrosis (CF), there has been a substantial effort to evaluate gene therapy as a modality for treating this disease. As the primary cause of mortality in CF is respiratory dysfunction, the l ...

AAV is a nonpathogenic human parvovirus that has a natural mechanism for long-term persistence in human cells. Wild-type AAV is unique in that it undergoes stable integration of its DNA into a specific region of human chromosome 19, the AAVS1 site (1–4), a process that our group has also recently descr ...

The amyloid precursor protein (APP) is a type I transmembrane protein with a large ectodomain, a single transmembrane domain and a small cytoplasmic tail (1). Translation of APP occurs at the endoplasmic reticulum (ER) and the protein is translocated into the ER lumen. The N-terminal domain of APP ...