Abstract: The N-degron is an intracellular degradation signal whose essential determinant is a specific, destabilizing, N-terminal amino acid residue. A set of N-degrons containing different destabilizing residues is manifested as the N-end rule, which relates the in vivo half-life of a protein to the identity of its N-terminal amino acid residue. Disclosed herein is a heat-inducible N-degron module. A heat-inducible N-degron module is a protein or peptide bearing a destabilizing N-terminal amino acid residue which becomes a substrate of the N-end rule pathway only at a temperature high enough to result in at least partial unfolding of the protein. At this elevated (nonpermissive) temperature, the heat-inducible N-degron module (and any protein or peptide attached at its C-terminus) is rapidly degraded in a cell in which the N-end rule pathway is operative.

Abstract: Disclosed is a pharmaceutical composition for oral delivery. The composition includes about 1-2 mg mammalian estrogen and about 25-100 mg phytoestrogen. Compositions of the type described above are utilized, for example, in a therapeutic regimen designed to reduce the risk of coronary heart disease and osteoporosis in postmenopausal women. This method comprises the oral administration of a composition comprising a mixture of estrogen and phytoestrogen, the dosages of mammalian estrogen and phytoestrogen being about 1-2 mg, about 25-100 mg, respectively.

Abstract: Methods of designing or modifying protein structure at the protein or genetic level to produce specified amino-termini in vivo or in vitro are described. The methods can be used to alter the metabolic stability and other properties of the protein or, alternatively, to artificially generate authentic amino-termini in proteins produced through artificial means. The methods are based upon the introduction of the use of artificial ubiquitin-protein fusions, and the discovery that the in vivo half-life of a protein is a function of the amino-terminal amino acid of the protein.

Abstract: The disclosure relates to a generic class of ubiquitin-specific proteases which specifically cleave at the C-terminus of the ubiquitin moiety in a ubiquitin fusion protein irrespective of the size of the ubiquitin fusion protein. More specifically, the disclosure relates to ubiquitin-specific proteases of this class which have been isolated from a cell. The disclosure also relates to isolated DNA sequences encoding the proteases of this class.

Abstract: Disclosed is a novel photo-activatable derivative of ryanodine 9-hydroxy-21-(4-azidobenzoyloxy)-9-epiryanodine. This novel ryanodine derivative has been conjugated to keyhole limpet hemocyanin (KLH) and used for the production of antibodies with high affinity and specificity to ryanodine. A radioimmunoassay specific for ryanodine was developed using the anti-ryanodine antibodies, and a dissociation constant for ryanodine of 1 nM was determined. Also disclosed is the binding of a labeled, photoactivatable derivative of ryanodine (ABRy) with the ryanodine receptor of skeletal, cardiac and brain membranes. Digestion of the labeled ryanodine receptor revealed a labeled 76 kDa tryptic fragment which is critical for the formation of the high affinity ryanodine binding site.

Abstract: The present invention relates to a method of diagnosing malignant hyperthermia susceptibility by detecting abnormal ryanodine receptor fragments following proteolytic enzyme digestion of ryanodine receptors isolated from the sarcoplasmic reticulum of mammalian skeletal muscle. The detection of abnormal ryanodine receptor fragments is indicative of malignant hyperthermia susceptibility.

Abstract: Disclosed are nucleic acid sequences encoding components of the dystrophin-glycoprotein complex. The components include dystroglycan, the 50 kDa protein component and the 59 kDa protein component. Also disclosed are compositions and methods which relate to the disclosed sequences.

Abstract: The invention pertains to pure, native dystrophin of mammalian skeletal muscle and a method of purifying dystrophin from mammalian skeletal muscle. The invention further pertains to a method of diagnosing muscular dystrophy by detecting the loss or abnormal structure of pure, native dystrophin from mammalian skeletal muscle. The detection of a loss of dystrophin or an abnormal structure of dystrophin is indicative of muscular dystrophy.

Abstract: Disclosed herein are methods for screening for primary cardiomyopathy. The methods are preferably immunological methods in which the level of binding of a monoclonal or polyclonal antibody to a 50 kD glycoprotein component of a mammalian muscle tissue is determined. This level of binding is compared to the level of binding observed when non-dystrophic tissue is treated in an otherwise identical manner. A substantial reduction in the level of binding to the 50 kD glycoprotein in the experimental mammalian muscle tissue has been determined to be a screen for primary cardiomyopathy.

Abstract: The invention pertains to the dystrophin-glycoprotein complex of mammalian skeletal muscle and a method of isolating said complex. The components of the complex and methods of separating and isolating said components also pertain to the invention. In addition, the invention further relates to a method of diagnosing muscular dystrophy by detecting and quantifying the loss of a non-dystrophin component of the dystrophin-glycoprotein complex with said loss being indicative of muscular dystrophy.

Abstract: A ubiquitin-specific protease which cleaves ubiquitin from any protein or peptide to which ubiquitin is joined and the gene encoding the protease are disclosed. The protease specifically cleaves the peptide bond in a fusion of ubiquitin to a protein or peptide between the carboxyl-terminal amino acid residue of a ubiquitin moiety and the .alpha.-amino group of any non-ubiquitin protein or peptide to which ubiquitin is joined. Recombinant expression vectors containing a DNA sequence encoding the ubiquitin-specific protease can be used to transform cells for production of the protease or to provide the cell with the ability to proteolytically deubiquitinate, in vivo, ubiquitin fusions co-produced by the cell. The protease can also be isolated and used to deubiquitinate ubiquitin fusions in vitro.

Abstract: Disclosed are methods for the diagnosis of autosomal muscular dystrophy through the analysis of muscle tissue using antibodies reactive with components of the dystrophin-glycoprotein complex. An experimental muscle tissue sample, treated if necessary to render components of the dystrophin-glycoprotein complex available for antibody binding, is contacted with an antibody which binds to a dystrophin-associated protein. The extent of antibody binding is determined, and compared to the extent of antibody binding to normal control tissue. A substantial reduction in the extent of binding to experimental tissue, as compared with normal control tissue, being diagnostic of autosomal muscular dystrophy. Among the autosomal muscular dystrophies which are detectable by the methods described herein are Fukuyama muscular dystrophy and severe childhood autosomal recessive muscular dystrophy.