232 results on '"Glenn E. Morris"'
Search Results
152. Diagnosis of X-linked Emery-Dreifuss muscular dystrophy by protein analysis of leucocytes and skin with monoclonal antibodies
- Author
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Caroline Sewry, Glenn E. Morris, Nguyen thi Man, Francesco Muntoni, and S. Manilal
- Subjects
Male ,X Chromosome ,Adolescent ,medicine.drug_class ,Genetic Linkage ,Emerin ,Fluorescent Antibody Technique ,Thymopoietins ,Biology ,Immunologic Tests ,Monoclonal antibody ,X-inactivation ,Muscular Dystrophies ,Western blot ,medicine ,Leukocytes ,Humans ,Muscular dystrophy ,Genetics (clinical) ,Skin ,medicine.diagnostic_test ,Genetic Carrier Screening ,Chromosome ,Antibodies, Monoclonal ,Membrane Proteins ,Nuclear Proteins ,medicine.disease ,Molecular biology ,Muscular Dystrophy, Emery-Dreifuss ,Xq28 ,Pedigree ,Neurology ,Pediatrics, Perinatology and Child Health ,Skin biopsy ,Female ,Neurology (clinical) - Abstract
The X-linked form of Emery-Dreifuss muscular dystrophy (EDMD) was recently shown to be due to mutations in the STA gene on chromosome Xq28. We have demonstrated a simple test for the diagnosis of this condition, looking for altered expression of the protein, emerin, in leucocytes and skin with a monoclonal antibody. Full-length emerin is completely absent in affected boys from the EDMD families studied. The method has also enabled identification of a female carrier of the disease by reduced levels of the protein on the leucocyte Western blot and a mosaic pattern of expression by immunofluorescence microscopy of the skin biopsy.
- Published
- 1997
153. P.5.18 Tissue-specific expression of nesprin isoforms and its relevance to muscular dystrophy and dilated cardiomyopathy
- Author
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Caroline Sewry, Glenn E. Morris, Ian Holt, D. Nguyen Thuy, Catherine M. Shanahan, L. Le Thanh Lam, and Qiuping Zhang
- Subjects
Gene isoform ,Nesprin ,Cardiac muscle ,Emerin ,Skeletal muscle ,Biology ,medicine.disease ,Embryonic stem cell ,Molecular biology ,medicine.anatomical_structure ,Neurology ,Pediatrics, Perinatology and Child Health ,medicine ,Neurology (clinical) ,Muscular dystrophy ,Genetics (clinical) ,Lamin - Abstract
A combination of isoform-specific, quantitative RT-PCR and site-specific monoclonal antibodies has been used to produce consistent evidence at both RNA and protein levels for the principal isoform products of SYNE1 and SYNE2, the genes encoding nesprin-1 and nesprin-2 proteins. Western blots show that both genes produce three groups of isoforms that are homologous in size and structure: giant, or full-length, isoforms of about 1000 kDa, medium isoforms of about 380 kDa (nesprin-1-beta and nesprin-2-gamma) and short isoforms of about 110 kDa (nesprin-1-alpha and nesprin-2-epsilon). SYNE2 alone produces a very short isoform at 60 kDa (nesprin-2-alpha) in skeletal muscle. Although the giant forms were the dominant isoforms in all tissues, major differences between tissues in expression of shorter isoforms were observed. From SYNE1, nesprin-1-alpha was found in skeletal and cardiac muscle only. Several cell lines, including embryonic stem cells, did not produce any detectable nesprin-1 at all. From SYNE2, we observed a developmental transition from nesprin-2-epsilon-1 at very early stages (embryonic stem cells, embryonal teratocarcinoma and ovary tissue) to nesprin-2-epsilon-2 in adult and fetal tissues. Both skeletal muscle and heart have a high proportion of short isoforms compared with other tissues, but heart expresses the nesprin-2-epsilon-2 isoform where skeletal muscle produces nesprin-2-alpha-1. This study has clarified the major nesprin isoforms expressed in cells and tissues, especially in cardiac and skeletal muscle, which are the major affected tissues in neuromuscular disease. Mutations in nesprin-1 and/or nesprin-2 can cause both Emery-Dreifuss muscular dystrophy and dilated cardiomyopathy. The operative functional unit appears to be the “LINC” complex of emerin, lamin A/C, nesprins and SUN proteins, since mutations in any single component can result in a similar pathology.
- Published
- 2013
154. Epitope Mapping Protocols
- Author
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Glenn E. Morris
- Subjects
Restriction enzyme ,Expression vector ,Epitope mapping ,Protein footprinting ,medicine.drug_class ,medicine ,Transposon mutagenesis ,Biology ,Monoclonal antibody ,Molecular biology ,Epitope ,Replacement method - Abstract
Overview: Choosing a Method for Epitope Mapping Glenn E. Morris Crystallographic Studies of Antigen-Antibody Interactions Frederick A. Saul and Pedro M. Alzari Epitope Mapping Antibody-Antigen Complexes by Nuclear Magnetic Resonance Spectroscopy Irina Kustanovich and Anat Zvi Mapping Epitopes on Antigens by Immunodiffusion in Gel Giuseppe A. Molinaro and William C. Eby A Simple Solid-Phase Competition Assay with Labeled Antigen Masahide Kuroki Epitope Mapping by Antibody Competition: Methodology and Evaluation of the Validity of the Technique Socrates J. Tzartos Epitope Mapping by Surface Plasmon Resonance in the BIAcore Berit Johne Identifying Residues in Antigenic Determinants by Chemical Modification N. Martin Young and Raymond P. Oomen Epitope Mapping by Differential Chemical Modification of Antigens Hans Rudolf Bosshard Epitope Mapping by Proteolysis of Antigen-Antibody Complexes: Protein Footprinting Ronald Jemmerson Proteolytic Fragmentation for Epitope Mapping Maria R. Mazzoni, Nickolai O. Artemyev, and Heidi E. Hamm Epitope Mapping by Chemical Fragmentation Glenn E. Morris Probing Antibody-Antigen Interactions by Mass Spectrometry Yingming Zhao and Brian T. Chait Epitope Mapping Using Multipin Peptide Synthesis Stuart J. Rodda, N. Joe Maeji, and Gordon Tribbick SPOT Synthesis: Epitope Analysis with Arrays of Synthetic Peptides Prepared on Cellulose Membranes Ronald Frank and Heike Overwin Tea Bag Synthesis of Positional Scanning Synthetic Combinatorial Libraries and Their Use for Mapping Antigenic Determinants Clemencia Pinilla, Jon R. Appel, and Richard A. Houghten Epitope Mapping Using Phage-Displayed Peptide Libraries Diane Dottavio Reiterative Screening of Phage-Display Peptide Libraries, with Antibodies Alexander Pereboev and Glenn E. Morris Homolog Scanning Lin-Fa Wang Epitope Mapping Using an Oligonucleotide Replacement Method Hannah Alexander Epitope Mapping by Region-Specified PCR Mutagenesis Takehiko Shibata and Masayuki Ikeda Random Fragment Libraries Using Yeast Expression Plasmid Serge Benichou and Genevieve Inchauspe Epitope Mapping Using Random Fragment Expression Libraries in l Phages Hartmut Porzig and Kenneth D. Philipson Random Fragment Libraries Displayed on Filamentous Phage Lin-Fa Wang and Meng Yu Epitope Mapping by Expression of Restriction Enzyme or PCR Fragments in Bacterial Plasmids Johannes A. Lenstra and Arnoud H. M. Van Vliet Epitope Mapping of Protein Antigens by Expression-PCR (E-PCR) David E. Lanar, Kevin C. Kain, and Henry B. Burch Epitope Mapping on Extracellular Domains of Cell-Surface Proteins Using Exonuclease III Thomas Brummendorf, Antonius Plagge, and Ullrich Treubert An Improved Method for Mapping Epitopes of Recombinant Antigens by Transposon Mutagenesis Steven G. Sedgwick, Brian A. Morgan, Nguyen thi Man, and Glenn E. Morris Incomplete Polypeptides of In Vitro Translation for Epitope Localization Lisa Djavadi-Ohaniance and Bertrand Friguet T-Cell Epitope Mapping with Synthetic Peptides and Peripheral Blood Mononuclear Cells Stuart J. Rodda Use of Natural or Selected Mutants and Variants for Epitope Mapping Glenn E. Morris Production of Panels of Monoclonal Antibodies by the Hybridoma Method Nguyen thi Man and Glenn E. Morris Production of Phage-Display Antibodies for Epitope Mapping Jenny Walker and George Banting Index
- Published
- 1996
155. The Emery-Dreifuss muscular dystrophy protein, emerin, is a nuclear membrane protein
- Author
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Nguyen thi Man, Caroline Sewry, Glenn E. Morris, and S. Manilal
- Subjects
Adult ,Recombinant Fusion Proteins ,Barrier-to-autointegration factor ,Emerin ,Thymopoietins ,Biology ,Muscular Dystrophies ,Fetus ,Genetics ,medicine ,Inner membrane ,Animals ,Humans ,Nuclear protein ,Emery–Dreifuss muscular dystrophy ,Molecular Biology ,Genetics (clinical) ,Cell Nucleus ,Muscles ,Antibodies, Monoclonal ,Membrane Proteins ,Nuclear Proteins ,General Medicine ,medicine.disease ,Molecular biology ,Cell nucleus ,medicine.anatomical_structure ,Epitope mapping ,Membrane protein ,Rabbits ,Epitope Mapping - Abstract
A large fragment of emerin cDNA was prepared by PCR and expressed as a recombinant protein in Escherichia coli. Using this as immunogen, we prepared a panel of 12 monoclonal antibodies which recognise at least four different epitopes on emerin in order to ensure that emerin can be distinguished from non-specific cross-reacting proteins. All the mAbs recognised a 34 kDa protein in all tissues tested, though minor emerin-related bands were also detected in some tissues. Immunofluorescence microscopy showed that emerin is located at the nuclear rim in all tissues examined. A muscle biopsy from an Emery-Dreifuss muscular dystrophy (EMDM) patient showed complete absence of emerin by both Western blotting and immunohistochemistry, suggesting a simple diagnostic antibody test for EDMD families. Biochemical fractionation of brain and liver tissues showed that emerin was present in nuclei purified by centrifugation through 65% sucrose and was absent from soluble fractions (post-100,000 g). From these results, together with sequence and structural homologies between emerin, thymopoietins and the nuclear lamina-associated protein, LAP2, we suggest that emerin will prove to be one member of a family of inner nuclear membrane proteins.
- Published
- 1996
156. Epitope Mapping of Protein Antigens by Competition ELISA
- Author
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Glenn E. Morris
- Subjects
Epitope mapping ,Antigen ,Linear epitope ,Chemistry ,media_common.quotation_subject ,Virology ,Competition (biology) ,Epitope ,media_common - Published
- 1996
157. Apo-dystrophins (Dp140 and Dp71) and dystrophin splicing isoforms in developing brain
- Author
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Glenn E. Morris, Nguyen thi Man, and C. Simmons
- Subjects
musculoskeletal diseases ,Gene isoform ,Adult ,Utrophin ,Duchenne muscular dystrophy ,Blotting, Western ,Molecular Sequence Data ,Biophysics ,Biology ,Biochemistry ,Polymerase Chain Reaction ,Dystrophin ,Exon ,medicine ,Humans ,RNA, Messenger ,Cloning, Molecular ,Molecular Biology ,Syntrophin ,Brain Chemistry ,Messenger RNA ,Base Sequence ,Antibodies, Monoclonal ,Brain ,Membrane Proteins ,Cell Biology ,Exons ,medicine.disease ,Molecular biology ,Alternative Splicing ,Cytoskeletal Proteins ,RNA splicing ,biology.protein - Abstract
PCR studies have shown that exons 71-74 are spliced out in most dystrophin mRNA transcripts in the brain. We have prepared new monoclonal antibodies against the syntrophin - binding region of dystrophin encoded by exons 73-74 and examined three protein products of the dystrophin gene in brain; the widely distributed Dp71, the recently discovered, brain-specific Dp140 and dystrophin itself. Exon 73-74 m Abs bound to all three proteins in brain and the extent of binding suggests that alternatively spliced dystrophins are less prominent at the protein level than predicted by PCR data. Dp140, unlike Dp71, was found to be present at much higher levels in foetal brain than in adult brain. If lack of functional Dp140 is the cause of the cognitive impairment in some Duchenne muscular dystrophy patients, this result suggests that the effects may occur early in development, which would reduce the options for therapeutic intervention.
- Published
- 1995
158. Characterization of genetic deletions in Becker muscular dystrophy using monoclonal antibodies against a deletion-prone region of dystrophin
- Author
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S. Hori, Victor Dubowitz, Le Thiet Thanh, Nguyen thi Man, Glenn E. Morris, and Caroline Sewry
- Subjects
musculoskeletal diseases ,medicine.drug_class ,Duchenne muscular dystrophy ,Molecular Sequence Data ,Gene mutation ,Monoclonal antibody ,Polymerase Chain Reaction ,Muscular Dystrophies ,Dystrophin ,Exon ,medicine ,Humans ,Amino Acid Sequence ,RNA, Messenger ,Muscular dystrophy ,Genetics (clinical) ,Southern blot ,Sequence Deletion ,Genetics ,biology ,Base Sequence ,Antibodies, Monoclonal ,medicine.disease ,Molecular biology ,Epitope mapping ,biology.protein ,Epitope Mapping ,Plasmids - Abstract
We have produced a new panel of 20 monoclonal antibodies (mAbs) against a region of the dystrophin protein corresponding to a deletion-prone region of the Duchenne muscular dystrophy gene (exons 45-50). We show that immuno-histochemistry or Western blotting with these "exon-specific" mAbs can provide a valuable addition to Southern blotting or PCR methods for the accurate identification of genetic deletions in Becker muscular dystrophy patients. The antibodies were mapped to the following exons: exon 45 (2 mAbs), exon 46 (6), exon 47 (1), exons 47/48 (4), exons 48-50 (6), and exon 50 (1). PCR amplification of single exons or groups of exons was used both to produce specific dystrophin immunogens and to map the mAbs obtained. PCR-mediated mutagenesis was also used to identify regions of dystrophin important for mAb binding. Because the mAbs can be used to characterize the dystrophin produced by individual muscle fibres, they will also be useful for studying "revertant" fibres in Duchenne muscle and for monitoring the results of myoblast therapy trials in MD patients with deletions in this region of the dystrophin gene.
- Published
- 1995
159. Evidence for a utrophin-glycoprotein complex in cultured cell lines and a possible role in cell adhesion
- Author
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Glenn E. Morris, Gareth E. Jones, Marian James, Clare J. Wise, and Catherine Simmons
- Subjects
Binding Sites ,Membrane Glycoproteins ,Utrophin ,Chemistry ,Macromolecular Substances ,Membrane Proteins ,Adhesion ,Biochemistry ,Actins ,Cell biology ,Cell Line ,Rats ,Dystrophin ,Cytoskeletal Proteins ,Mice ,Glycoprotein complex ,Cultured cell ,Cell Adhesion ,Animals ,Humans ,Neural cell adhesion molecule ,Cell adhesion ,Dystroglycans ,Glycoproteins - Published
- 1995
160. Epitope mapping of recombinant antigens by transposon mutagenesis
- Author
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Glenn E. Morris, Steven G. Sedgwick, and Nguyen thi Man
- Subjects
Transposable element ,Blotting, Western ,Molecular Sequence Data ,Reading frame ,Bioengineering ,Biology ,Applied Microbiology and Biotechnology ,Biochemistry ,Epitope ,Dystrophin ,Plasmid ,Escherichia coli ,Antigens ,Molecular Biology ,Genetics ,Base Sequence ,Antibodies, Monoclonal ,Sequence Analysis, DNA ,Sleeping Beauty transposon system ,Recombinant Proteins ,Subcloning ,Epitope mapping ,Mutagenesis ,DNA Transposable Elements ,Transposon mutagenesis ,Epitope Mapping ,Biotechnology - Abstract
We describe a method for generating a plasmid library expressing random truncations of a recombinant protein and for epitope mapping by screening the library with monoclonal antibodies. The key step is the random introduction of the transposon, Tn1000, which carries stop codons in all three reading frames, into a bacterial expression plasmid by using a simple bacterial mating procedure. Antibody-positive clones are then selected and the point of protein truncation is determined by sequencing the plasmid DNA at the point of transposon insertion. One advantage of the method is that no subcloning or in vitro manipulation of DNA is necessary.
- Published
- 1995
161. The N-terminal half of dystrophin is protected from proteolysis in situ
- Author
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Glenn E. Morris, Nguyen thi Man, S. Ohtani, and S. Hori
- Subjects
musculoskeletal diseases ,In situ ,congenital, hereditary, and neonatal diseases and abnormalities ,Proteases ,medicine.drug_class ,Dystroglycan binding ,Proteolysis ,Blotting, Western ,Biophysics ,Monoclonal antibody ,Biochemistry ,Dystrophin ,Endopeptidases ,medicine ,Humans ,Cytoskeleton ,Muscle, Skeletal ,Molecular Biology ,Binding Sites ,biology ,medicine.diagnostic_test ,Chemistry ,Antibodies, Monoclonal ,Cell Biology ,Exons ,musculoskeletal system ,Molecular biology ,Protein tertiary structure ,Protein Structure, Tertiary ,Models, Structural ,Postmortem Changes ,biology.protein - Abstract
Using a panel of "exon-specific" monoclonal antibodies, we have examined the products of degradation of dystrophin by endogenous proteases in post-mortem human muscle. Four main sites of dystrophin digestion were identified, all of them in the C-terminal half of the molecule. Two of them correspond to "hinges" in the central rod region and a third in the C-terminal domain follows the dystroglycan binding site. The results support the Koenig and Kunkel model for the tertiary structure of dystrophin (J. Biol. Chem. 265 (1990) 4560-4566), but suggest that much of the N-terminal half of dystrophin is protected from proteolysis, possibly by interaction with the sub-sarcolemmal cytoskeleton. Although the results seem inconsistent with an anti-parallel dimer model of dystrophin in which hinge 2 and hinge 3 are close together, possible ways of reconciling them with such a model are also considered.
- Published
- 1995
162. Full-length and short forms of utrophin, the dystrophin-related protein
- Author
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Timothy R. Helliwell, John Kendrick-Jones, Kay E. Davies, C. Simmons, Nguyen thi Man, Glenn E. Morris, and Steven J. Winder
- Subjects
Monoclonal antibody ,Actin binding ,Utrophin ,medicine.drug_class ,animal diseases ,Biophysics ,Neuromuscular junction ,Biochemistry ,Dermatomyositis ,Dystrophin ,Mice ,Structural Biology ,Genetics ,medicine ,Tumor Cells, Cultured ,Animals ,Humans ,Muscular dystrophy ,Molecular Biology ,Mice, Inbred BALB C ,Sarcolemma ,biology ,Antibodies, Monoclonal ,Membrane Proteins ,Cell Biology ,medicine.disease ,musculoskeletal system ,Molecular biology ,Immunohistochemistry ,Blot ,Cytoskeletal Proteins ,medicine.anatomical_structure ,biology.protein ,Antibody - Abstract
All previous studies of the localization of utrophin (the dystrophin-related protein) in muscle and other tissues have been performed only with antibodies against the C-terminal region of the protein. Since several short forms of dystrophin, the apodystrophins, are produced from the 3′ end of the dystrophin gene, there is a possibility that similar short forms of utrophin exist and that these could be responsible for some of the many different localizations of ‘utrophin’ in muscle. We have produced a new panel of 15 mAbs against the N-terminal region of utrophin and we have used it together with mAbs against the C-terminal region to show that full-length utrophin is present at neuromuscular junctions, in nerves, blood vessels and capillaries in normal muscle and in the sarcolemma of patients with muscular dystrophy and dermatomyositis. However, two of the 15 mAbs also recognised rat/mouse utrophin and both of these detected an additional 62 kDa protein on Western blots of rat C6 glioma cells. This potential 62 kDa ‘apo-utrophin’ was not detected in human cerebral cortex, in rat Schwannoma cells nor in any of the non-nerve cells and tissues tested.
- Published
- 1995
163. Screening donor blood for malaria by polymerase chain reaction
- Author
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E. O'Brien, Luong Van Hien, Tran van Be, Phan Ngoc Tran, Vu thi Ty Hang, Le Thiet Thanh, and Glenn E. Morris
- Subjects
Blood transfusion ,medicine.medical_treatment ,Plasmodium vivax ,Molecular Sequence Data ,Plasmodium falciparum ,Blood Donors ,Parasitemia ,Polymerase Chain Reaction ,Sensitivity and Specificity ,law.invention ,law ,parasitic diseases ,medicine ,Malaria, Vivax ,Animals ,Humans ,Malaria, Falciparum ,Polymerase chain reaction ,biology ,Base Sequence ,Transmission (medicine) ,business.industry ,Viet nam ,Public Health, Environmental and Occupational Health ,Transfusion Reaction ,General Medicine ,biology.organism_classification ,medicine.disease ,Virology ,Ho chi minh ,Infectious Diseases ,Immunology ,Parasitology ,business ,Malaria - Abstract
In countries where malaria is endemic, its transmission is a hazard of blood transfusion. The microscopical and immunological methods in current use for malaria diagnosis are unsatisfactory for low levels of parasitaemia in blood donations. The polymerase chain reaction (PCR) can be 100-fold more sensitive than thick blood film examination when appropriate primers are used and can detect and distinguish Plasmodium falciparum and P. vivax in a single tube. A study of 1506 blood donations in Ho Chi Minh City (3 of which were positive) suggests that PCR can provide an effective screen for P. falciparum under local conditions. Studies in a region of Viet Nam where malaria is common showed that PCR detects many more cases of low-level parasitaemia (19/30) than thick blood films (4/30).
- Published
- 1995
164. Changes at the N-terminus of human brain creatine kinase during a transition between inactive folding intermediate and active enzyme
- Author
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Glenn E. Morris and Nguyen thi Man
- Subjects
Protein Conformation ,Molecular Sequence Data ,Biophysics ,Peptide ,Biochemistry ,Epitope ,chemistry.chemical_compound ,Epitopes ,Structure-Activity Relationship ,Protein structure ,Structural Biology ,Peptide synthesis ,Humans ,Amino Acid Sequence ,Molecular Biology ,Creatine Kinase ,chemistry.chemical_classification ,biology ,Antibodies, Monoclonal ,Brain ,Peptide Fragments ,Amino acid ,Epitope mapping ,chemistry ,biology.protein ,Creatine kinase ,Protein folding - Abstract
CK-STAR, a monoclonal antibody against human brain creatine kinase (CK), can be shown by chemical cleavage mapping and peptide synthesis to recognize an epitope at the free N-terminus of the enzyme. The epitope could be largely reproduced by a synthetic peptide based on the first 18 amino acids and could be partly formed by the first 11 amino acids. The antibody did not bind to native CK, but it did bind to CK in various partially denatured forms and to an enzymically inactive intermediate in the refolding process. Competitive binding studies have shown that the N-terminal conformations of both the refolding intermediate and the free peptide resemble that of CK partially denatured by attachment to plastic. The results suggest that the final stages of CK refolding and reactivation involve a structural change at the N-terminus or its interaction with some other part of the CK molecule, thus masking the CK-STAR epitope.
- Published
- 1992
165. Book reviews
- Author
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David B. Guiliano, Ruth McNerney, Glenn E. Morris, and Peter Shepherd
- Subjects
Bioengineering ,Molecular Biology ,Applied Microbiology and Biotechnology ,Biochemistry ,Biotechnology - Published
- 2000
166. Structural relationships between hepatitis B surface antigen in human plasma and dimers from recombinant vaccine: a monoclonal antibody study
- Author
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Phan Ngoc Tran, Buu Mat, Nguyen thi Vinh Ha, Glenn E. Morris, Nguyen thi Man, and Le Thiet Thanh
- Subjects
Cancer Research ,HBsAg ,Sucrose ,medicine.drug_class ,Blotting, Western ,Enzyme-Linked Immunosorbent Assay ,Monoclonal antibody ,Binding, Competitive ,Epitope ,law.invention ,Epitopes ,Mice ,Structure-Activity Relationship ,Viral envelope ,Antigen ,law ,Antibody Specificity ,Virology ,medicine ,Centrifugation, Density Gradient ,Animals ,Humans ,Mice, Inbred BALB C ,Vaccines, Synthetic ,Hepatitis B Surface Antigens ,biology ,Antibodies, Monoclonal ,Infant ,biology.organism_classification ,Molecular biology ,Infectious Diseases ,Hepadnaviridae ,biology.protein ,Recombinant DNA ,Electrophoresis, Polyacrylamide Gel ,Antibody - Abstract
Ten monoclonal antibodies were obtained from mice immunized with a yeast recombinant hepatitis B vaccine. They were selected at an early stage for their ability to bind to native surface antigen particles (HBsAg) in human plasma. All antibodies recognized conformational epitopes which were destroyed completely or almost completely by reduction of disulphide bridges. They were divided into five epitope groups by their competition for binding to recombinant S protein, though epitopes within each group are not identical. Recombinant S protein migrated on SDS-PAGE in the absence of reducing agents as a mixture of monomers and dimers/oligomers. Sucrose gradient analysis suggests that all these forms are co-aggregated into HBsAg-Iike particles. On Western blots, all ten antibodies either bound only to dimers/oligomers or strongly preferred them over monomers. The results suggest that, of the antibodies produced in response to recombinant vaccine in mice, most of those which bind strongly to ‘native’ HBsAg particles in human plasma recognize surface structures created by interaction between two subunits.
- Published
- 1991
167. Fetal dystrophin to diagnose carrier status
- Author
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Antoon F.M. Moorman, A. van Haeringen, I. B. Ginjaar, G.J.B. van Ommen, Glenn E. Morris, L V Nicholson, S. Soffers, Egbert Bakker, and Other departments
- Subjects
Male ,Heterozygote ,Fetus ,Pathology ,medicine.medical_specialty ,biology ,business.industry ,Duchenne muscular dystrophy ,DNA ,General Medicine ,medicine.disease ,Muscular Dystrophies ,Pedigree ,Dystrophin ,Fetal Diseases ,medicine ,biology.protein ,Carrier status ,Humans ,Female ,business - Published
- 1991
168. D.P.4.10 Muscleblind-like proteins: Similarities and differences in normal and myotonic dystrophy muscle
- Author
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V. Jacquemin, Majid Fardaei, Ian Holt, Gillian Butler-Browne, Glenn E. Morris, J.D. Brook, Caroline Sewry, and Denis Furling
- Subjects
medicine.medical_specialty ,Endocrinology ,Neurology ,Internal medicine ,Pediatrics, Perinatology and Child Health ,medicine ,Neurology (clinical) ,Biology ,medicine.disease ,Myotonic dystrophy ,Genetics (clinical) - Published
- 2008
169. Monoclonal antibody studies suggest a catalytic site at the interface between domains in creatine kinase
- Author
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Alison J. Cartwright and Glenn E. Morris
- Subjects
medicine.drug_class ,Molecular Sequence Data ,Biophysics ,Monoclonal antibody ,Biochemistry ,Epitope ,Conserved sequence ,Epitopes ,Structural Biology ,Sequence Homology, Nucleic Acid ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Cyanogen Bromide ,Binding site ,Molecular Biology ,Creatine Kinase ,Binding Sites ,biology ,Muscles ,Myocardium ,Active site ,Antibodies, Monoclonal ,Molecular biology ,Peptide Fragments ,Isoenzymes ,Epitope mapping ,biology.protein ,Protein folding ,Creatine kinase ,Chickens - Abstract
We have located the epitopes recognized by four different monoclonal antibodies which bind to partially unfolded creatine kinase (CK) (ATP: creatine N-phosphotransferase, EC 2.7.3.2) but not to the native enzyme. The epitopes appear to be buried within the CK structure in its native, proteinase-resistant, state. When the epitopes are made accessible to antibody by mild denaturation, CK becomes enzymically-inactive and can be cleaved by proteinase V8 into two large fragments which retain the epitopes and may represent domains. Epitopes on each V8 fragment are associated with highly conserved sequences and are brought physically close to the active site of the enzyme during the later stages of CK refolding and reactivation. The results suggest a catalytic site formed at the interface between two domains which carry the epitopes on their interacting surfaces. Separation of loosely associated domains before or during immunization may account for the origin of antibodies against buried epitopes.
- Published
- 1990
170. Specificity of dystrophin analysis improved with monoclonal antibodies
- Author
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Nguyen thi Man, Glenn E. Morris, I. B. Ginjaar, G.J.B. van Ommen, Antoon F.M. Moorman, J.M. Ellis, and Other departments
- Subjects
Pathology ,medicine.medical_specialty ,Staining and Labeling ,biology ,medicine.drug_class ,Carrier state ,Diagnostico diferencial ,Antibodies, Monoclonal ,Dystrophy ,General Medicine ,Monoclonal antibody ,Muscular Dystrophies ,Diagnosis, Differential ,Dystrophin ,Antibodies monoclonal ,Carrier State ,biology.protein ,medicine ,Humans ,Differential diagnosis ,Antibody - Published
- 1990
171. Is myoblast transplantation effective?
- Author
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Qi Long Lu, Glenn E. Morris, T. Partridge, and Eric P. Hoffman
- Subjects
Transplantation ,Genetics ,General Medicine ,Computational biology ,Biology ,General Biochemistry, Genetics and Molecular Biology - Published
- 1998
172. M.P.4.06 Model systems for developing therapies for McArdle disease
- Author
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Caroline Sewry, K. Wright, Glenn E. Morris, and Ros Quinlivan
- Subjects
Oncology ,medicine.medical_specialty ,MCARDLE DISEASE ,Neurology ,business.industry ,Internal medicine ,Pediatrics, Perinatology and Child Health ,medicine ,Neurology (clinical) ,business ,Genetics (clinical) - Published
- 2007
173. Early onset X-linked Emery-Dreifuss muscular dystrophy resembling limb-girdle muscular dystrophy
- Author
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E.J. Lichtarowicz-Krynska, Francesco Muntoni, Victor Dubowitz, D. Recan, Stéphane Llense, Caroline Sewry, J.-C. Kaplan, Glenn E. Morris, Jeremy M. G. Taylor, and S. Manilal
- Subjects
Neurology ,business.industry ,Pediatrics, Perinatology and Child Health ,X-Linked Emery-Dreifuss Muscular Dystrophy ,medicine ,Neurology (clinical) ,Anatomy ,medicine.disease ,business ,Genetics (clinical) ,Early onset ,Limb-girdle muscular dystrophy - Published
- 1997
174. Monitoring of recombinant survival motor neuron protein using fiber-optic surface plasmon resonance
- Author
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Christian L. Lorson, Philip J. Young, Margaret Barnhart, Tina M. Battaglia, Karl S. Booksh, Glenn E. Morris, Jean-Francois Masson, and Ronald A. Nieman
- Subjects
animal diseases ,Nerve Tissue Proteins ,Biosensing Techniques ,SMN1 ,Biochemistry ,Analytical Chemistry ,law.invention ,chemistry.chemical_compound ,law ,Electrochemistry ,medicine ,Fiber Optic Technology ,Humans ,Environmental Chemistry ,Surface plasmon resonance ,Optical Fibers ,Spectroscopy ,Motor Neurons ,medicine.diagnostic_test ,Chemistry ,Spinal muscular atrophy ,Surface Plasmon Resonance ,Motor neuron ,medicine.disease ,SMA ,Molecular biology ,Recombinant Proteins ,nervous system diseases ,medicine.anatomical_structure ,nervous system ,Myoglobin ,Immunoassay ,Recombinant DNA - Abstract
Spinal muscular atrophy (SMA) is the leading genetic cause of infant mortality. SMA is caused by the homozygous loss of the survival motor neuron 1 (SMN1) gene. A nearly identical copy gene exists known as SMN2, however, due to an aberrant splicing event, the SMN2 gene fails to produce sufficient full-length protein to protect against disease development in the absence of SMN1. While a number of compounds have recently been identified that can stimulate full-length survival motor neuron (SMN) expression from the nearly identical copy SMN2, one of the difficulties has been the lack of a highly reproducible and quantitative means to measure the levels of SMN protein. To develop a technique that allows the rapid and highly sensitive measurement of SMN protein, a Surface Plasmon Resonance (SPR) application has been developed. The ability to quantify unassociated SMN protein and monitor the binding of SMN with other proteins in solution using a SPR sensor in less than 15 min and at low ng mL(-1) levels in HEPES Buffer Saline (HBS) has been achieved. The detection limit for the specific binding of SMN in HBS pH 7.4 solution is 0.99 ng mL(-1) with non-specific binding accounting for approximately 30% of the signal. Quantification of SMN is based on an immunoassay performed on the gold surface of the SPR sensor. 16-mercaptohexadecanoic acid (MHA) was reacted with dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) to form a pre-activated thiol (MHA-NHS). Antibodies for SMN were then coupled to the sensor with the pre-activated thiol. Sensor specificity was examined with mixtures of myoglobin (MG) and SMN. SMN sensor response decreases by more than 60% when MG was added to SMN. The decrease in sensor response can be attributed to non-specific binding of SMN to MG, verified with a sensor for MG.
- Published
- 2004
175. Problems with LAP nomenclature
- Author
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Robert Craigie, Roland Foisner, Henning Otto, Christopher J. Hutchison, Brian Burke, Glenn E. Morris, Yosef Gruenbaum, Amos J. Simon, Ricardo Benavente, Crafford A. Harris, Georg Krohne, Kazuhiro Furukawa, Larry Gerace, Katherine L. Wilson, Robert D. Goldman, and Howard J. Worman
- Subjects
Computer science ,Cell Biology ,Computational biology ,Nomenclature ,Cell biology - Published
- 2001
176. The molecular pathogenesis of a disease-causing mutation (g993t) in the emerin gene: an in vitro mutagenesis study
- Author
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S. Manual, I. Holt, L. Clements, and Glenn E. Morris
- Subjects
Genetics ,Molecular pathogenesis ,Emerin ,Mutagenesis (molecular biology technique) ,Disease-causing Mutation ,Biology ,Biochemistry ,Gene ,In vitro - Published
- 2000
177. Household Behavior under Alternative Pay-as-You-Throw Systems for Solid Waste Disposal
- Author
-
George L. Van Houtven and Glenn E. Morris
- Subjects
Economics and Econometrics ,Environmental Science (miscellaneous) - Published
- 1999
178. Disruption of the utrophin‒actin interaction by monoclonal antibodies and prediction of an actin-binding surface of utrophin
- Author
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Glenn E. MORRIS, Nguyen thi MAN, Nguyen thi Ngoc HUYEN, Alexander PEREBOEV, John KENDRICK-JONES, and Steven J. WINDER
- Subjects
Cell Biology ,Molecular Biology ,Biochemistry - Published
- 1999
179. Full length huntingtin is not detected in intranuclear inclusions in Huntington's disease brain
- Author
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James Neal, T M Nguyen, Fiona L. Wilkinson, Peter S. Harper, A L Jones, Glenn E. Morris, and Philip Thomas
- Subjects
Cell Nucleus ,Cerebral Cortex ,Inclusion Bodies ,Huntingtin Protein ,Pathology ,medicine.medical_specialty ,Huntingtin ,Chemistry ,Intranuclear Inclusions ,Nuclear Proteins ,Nerve Tissue Proteins ,medicine.disease ,Immunohistochemistry ,Biochemistry ,Huntington Disease ,Huntington's disease ,medicine ,Humans ,Caudate Nucleus - Published
- 1998
180. A family with severe pseudo-dominant Emery-Dreifuss muscular dystrophy due to emerin deficiency
- Author
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F. Leturcq, J. Bensaid, Jon Andoni Urtizberea, D. Recan, Glenn E. Morris, F. Fraisse, P. Warrot, Stéphane Llense, J.-C. Kaplan, N. Deburgrave, J.-M. Dupont, D. Amsallem, C. Giraudet, and J.-C. Barbot
- Subjects
Pathology ,medicine.medical_specialty ,Neurology ,business.industry ,Pediatrics, Perinatology and Child Health ,Emerin ,Medicine ,Neurology (clinical) ,Emery–Dreifuss muscular dystrophy ,business ,medicine.disease ,Genetics (clinical) - Published
- 1997
181. X-linked Emery-Dreifuss muscular dystrophy: molecular diagnosis by protein analysis and use of the skin biopsy in female carriers
- Author
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Glenn E. Morris, J. Pradas, S. Manilal, M. Guitet, Jaume Colomer, J. Corbera, and J. Vila
- Subjects
Pathology ,medicine.medical_specialty ,Neurology ,medicine.diagnostic_test ,business.industry ,Pediatrics, Perinatology and Child Health ,Skin biopsy ,X-Linked Emery-Dreifuss Muscular Dystrophy ,medicine ,Neurology (clinical) ,business ,Genetics (clinical) - Published
- 1997
182. Exon-specific monoclonal antibodies against dystrophn for diagnosis and genetic deletion analysis in muscular dystrophy
- Author
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Le Thiet Thanh, Glenn E. Morris, and Nguyen thi Man
- Subjects
Exon ,Neurology ,medicine.drug_class ,business.industry ,Pediatrics, Perinatology and Child Health ,medicine ,Neurology (clinical) ,Muscular dystrophy ,Monoclonal antibody ,business ,medicine.disease ,Molecular biology ,Genetics (clinical) - Published
- 1996
183. Monoclonal antibodies against emerin, for the diagnosis of emery-dreifuss muscular dystrophy
- Author
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Glenn E. Morris, Sushila Maniw, Nguyen thi Man, and Caroline Sewry
- Subjects
Pathology ,medicine.medical_specialty ,business.industry ,medicine.drug_class ,Emerin ,medicine.disease ,Monoclonal antibody ,Neurology ,Pediatrics, Perinatology and Child Health ,medicine ,Neurology (clinical) ,Emery–Dreifuss muscular dystrophy ,business ,Genetics (clinical) - Published
- 1996
184. Probing protein structure with proteases: studies of an equilibrium intermediate in protein unfolding
- Author
-
Timothy I. Webb, Philip J. Jackson, and Glenn E. Morris
- Subjects
Protein Folding ,Proteases ,Molecular Structure ,Sequence Homology, Amino Acid ,Protein Conformation ,Chemistry ,Molecular Sequence Data ,Proteins ,Arginine Kinase ,Biochemistry ,Protein Structure, Secondary ,Nephropidae ,Crystallography ,Protein structure ,Endopeptidases ,Biophysics ,Unfolded protein response ,Animals ,Amino Acid Sequence ,Chickens ,Creatine Kinase - Published
- 1995
185. Monitoring of recombinant survival motor neuron protein using fiber-optic surface plasmon resonance.
- Author
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Jean-Francois Masson, Margaret Barnhart, Tina M. Battaglia, Glenn E. Morris, Ronald A. Nieman, Philip J. Young, Christian L. Lorson, and Karl S. Booksh
- Published
- 2004
186. Asbestos fibre supply and the regulation of asbestos exposure
- Author
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Glenn E. Morris and Stephen E. Margolis
- Subjects
Economics and Econometrics ,Sociology and Political Science ,Asbestos fibre ,Natural resource economics ,medicine ,Business ,Management, Monitoring, Policy and Law ,medicine.disease_cause ,Law ,Domestic market ,Asbestos - Abstract
The USA and Canada are each considering measures to reduce general exposure to asbestos. Product bans and workplace exposure rules may each reduce general exposure by reducing domestic asbestos demand. The effectiveness of demand-reducing measures will depend upon the elasticity of asbestos supply to the domestic market. This paper provides calculations of this elasticity for three fibre grade groupings, using a framework that recognizes international trade in asbestos and the joint product nature of most asbestos fibre production. The results suggest that reductions in North American demand result in relatively substantial changes in asbestos use but very small changes in price.
- Published
- 1986
187. Specificity of Nuclear Protein Kinases in Differentiating Chick Skeletal-Muscle Cells
- Author
-
Nguyen thi Man, R.J. Cole, and Glenn E. Morris
- Subjects
medicine.anatomical_structure ,Chemistry ,Kinase ,medicine ,Skeletal muscle ,Nuclear protein ,Biochemistry ,Cell biology - Published
- 1978
188. Monoclonal antibody studies of creatine kinase. The ART epitope: evidence for an intermediate in protein folding
- Author
-
Glenn E. Morris
- Subjects
Protein Conformation ,Molecular Sequence Data ,Biochemistry ,Epitope ,Epitopes ,chemistry.chemical_compound ,Animals ,Urea ,Trypsin ,Amino Acid Sequence ,Cysteine ,Amino Acids ,Creatine Kinase ,Molecular Biology ,Gel electrophoresis ,chemistry.chemical_classification ,Methionine ,biology ,Antibodies, Monoclonal ,Cell Biology ,Proteinase K ,Molecular biology ,Amino acid ,chemistry ,biology.protein ,Creatine kinase ,Rabbits ,Chickens ,Research Article ,Conformational epitope - Abstract
Chemical cleavage at cysteine residues with nitrothiocyanobenzoic acid shows that the last 98 amino acids of the 380-amino-acid sequence of chick muscle creatine kinase are sufficient for binding of the monoclonal antibody CK-ART. Removal of the last 30 amino acids by cleavage at methionine residues with CNBr results in loss of CK-ART binding. CK-ART binding is also lost when these C-terminal methionine residues are oxidized to sulphoxide, but binding is regained on reduction. Proteinase K ‘nicks’ native CK at a single site near the C-terminus and two fragments of 327 amino acides and 53 amino acids can be separated by subsequent SDS or urea treatment. CK-ART still binds normally to ‘nicked’ CK, which is enzymically inactive. After treatment with either urea (in a competition enzyme-linked immunosorbent assay) or SDS (on Western blots), however, CK-ART binds to neither of the two fragments, although these treatments do not affect binding to intact CK. This suggests that parts of both CK fragments contribute to the CK-ART epitope. CK-ART is both species- and isoenzyme-specific, binding only to chick M-CK. The only C-terminal regions containing chick-specific sequences are residues 300-312 and residues 368-371, the latter group being close to the essential methionine residues. We suggest that one, or possibly both, of these regions is involved in forming the conformational epitope on the surface of the CK molecule which CK-ART recognizes. Native CK is resistant to trypsin digestion. The C-terminal half of urea-treated and partly-refolded CK is also resistant to trypsin digestion, whereas the N-terminal half is readily digested. The results suggest a C-terminal region which can refold more rapidly than the rest of the CK molecule and provide evidence for an intermediate in CK refolding.
- Published
- 1989
189. Two-dimensional gel analysis of nuclear proteins during muscle differentiation in vitro
- Author
-
Nguyen thi Man, R.J. Cole, and Glenn E. Morris
- Subjects
Gel electrophoresis ,Lysis ,Cellular differentiation ,Phosphatase ,Cell Biology ,Biology ,Molecular biology ,Tropomyosin ,In vitro ,humanities ,chemistry.chemical_compound ,Tubulin ,chemistry ,Biochemistry ,Cytoplasm ,biology.protein ,Myocyte ,Phosphorylation ,Protein phosphorylation ,Nuclear protein ,Protein kinase A ,Actin ,DNA - Abstract
Isolated nuclei from chick skeletal muscle cell cultures at different stages of differentiation were labelled in vitro with γ-[ 32 P]ATP. Nuclear proteins were separated into saline-soluble and non-histone (NHP) fractions and analysed by two-dimensional gel electrophoresis and autoradiography. Relatively few proteins (10–12) were phosphorylated to any extent and most of these corresponded to minor, or even undetectable, components on stained gels. 32 P incorporation into most of these proteins increased between 24 and 40 h and decreased sharply between 40 and 66 h, the changes being more marked in some phosphoproteins than in others. Pulse-chase data suggest that these overall changes in phosphorylation reflect changes in protein kinase, rather than protein phosphatase activities. One highly phosphorylated acidic protein (mol. wt 10 000 D; pI 4.2) which was undetectable on stained gels, showed a continuous increase in 32 P incorporation between 24 and 66 h in both saline-soluble and NHP fractions. This protein may be the muscle counterpart of a phosphorylated nuclear protein partially characterized in other cell types. Both nuclear protein phosphorylation and low molecular weight non-histone proteins have been particularly implicated in activation of gene transcription in other systems, although we have no direct evidence, as yet, to connect them with control of myoblast growth and differentiation.
- Published
- 1980
190. A mechanical dissociation method for preparation of muscle cell cultures
- Author
-
Glenn E. Morris, Katherine Tepperman, Stuart M. Heywood, and Francine Essien
- Subjects
Cell Survival ,Physiology ,Muscles ,Cell preparation ,Clinical Biochemistry ,Skeletal muscle ,Cell Separation ,Cell Biology ,Anatomy ,Biology ,Trypsin ,Suspension culture ,Dissociation (chemistry) ,Trypsinization ,Cell biology ,medicine.anatomical_structure ,Polyribosomes ,medicine ,Myocyte ,Embryonic chick ,Cells, Cultured ,medicine.drug - Abstract
A cell preparation method by which large numbers of embryonic chick skeletal muscle cells may be obtained is described. The procedure requires fewer manipulations and much less time than standard trypsinization. By the criteria used, both methods are comparable with respect to percent viable cells and survival of plated cells. However, in addition to the ease of preparation, the mechanical dissociation method offers the significant advantage that the cell suspension is greatly enriched for myoblasts without the necessity of an additional preplating step.
- Published
- 1975
191. Treatment of human muscle creatine kinase with glutaraldehyde preferentially increases the immunogenicity of the native conformation and permits production of high-affinity monoclonal antibodies which recognize two distinct surface epitopes
- Author
-
Alison J. Cartwright, Glenn E. Morris, Nguyen thi Man, and Karen M. Andrews
- Subjects
biology ,Protein Conformation ,medicine.drug_class ,Immunogenicity ,Immunology ,Antibody Affinity ,Antibodies, Monoclonal ,Monoclonal antibody ,Molecular biology ,Epitope ,Epitopes ,Epitope mapping ,Antigen ,Biochemistry ,Antibody Specificity ,Glutaral ,biology.protein ,medicine ,Native state ,Humans ,Immunology and Allergy ,Creatine kinase ,Antibody ,Creatine Kinase - Abstract
Treatment of human muscle creatine kinase (MM-CK) with glutaraldehyde produced highly aggregated forms which retained the native antigenic structure. Immunization of BALB/c mice with CK aggregates instead of untreated CK produced over ten-fold higher titres of antibody against native CK without increasing the tires of antibody against denatured enzyme. Production of high-affinity monoclonal antibodies specific for both the muscle isoenzyme and the native conformation became possible where the use of untreated CK had failed. Four monoclonal antibodies have been characterized by an epitope mapping technique and compared with a commercially available monoclonal antibody. One antibody has a much higher affinity for MM-CK than the other three and the commercial antibody. Competition studies show that it also recognizes a different epitope on the CK surface from the other three monoclonal antibodies which bind to the same surface region as the commercial antibody. Immunoassays based on the high affinity antibody can easily measure less than 1 ng of CK, a sensitivity comparable to, or better than, standard enzymatic assays.
- Published
- 1989
192. Monoclonal-antibody studies of creatine kinase. The proteinase K-cleavage site
- Author
-
L P Head, Glenn E. Morris, and L C Frost
- Subjects
Models, Molecular ,Protein Conformation ,Peptide ,Cleavage (embryo) ,Biochemistry ,Proteinase 3 ,Endopeptidases ,Amino Acid Sequence ,Creatine Kinase ,Immunoelectrophoresis ,Molecular Biology ,Peptide sequence ,Protein secondary structure ,chemistry.chemical_classification ,Binding Sites ,biology ,Antibodies, Monoclonal ,Cell Biology ,Proteinase K ,Molecular biology ,Peptide Fragments ,Enzyme ,chemistry ,biology.protein ,Electrophoresis, Polyacrylamide Gel ,Creatine kinase ,Endopeptidase K ,Research Article - Abstract
Proteinase K cleaves a small peptide from native muscle-specific creatine kinase. We present evidence, from the binding of two monoclonal antibodies to formic acid-cleavage fragments and proteinase K-digest fragments of chick muscle creatine kinase, that the proteinase K-cleavage site is in the C-terminal region of the molecule. This specificity of proteinase K, which is not normally a highly specific enzyme, and the continued association of the two peptide fragments after cleavage suggest an unusual conformational feature in the cleavage-site region. By applying predictive methods for hydrophobicity and secondary structure to an amino acid sequence in this region, we suggest possible structural features at the cleavage site that are evidently conserved across avian and mammalian species. The most likely site is next to, or within, a beta-turn on the surface of the molecule.
- Published
- 1985
193. Biosynthesis of muscle-specific creatine kinase during differentiation in vitro
- Author
-
Glenn E. Morris and R.J. Cole
- Subjects
biology ,Chemistry ,Muscles ,Biophysics ,Cell Differentiation ,Chick Embryo ,Cell Biology ,Mitogen-activated protein kinase kinase ,Biochemistry ,In vitro ,Isoenzymes ,chemistry.chemical_compound ,P70S6 kinase ,Biosynthesis ,Structural Biology ,Genetics ,biology.protein ,Animals ,Creatine kinase ,Creatine Kinase ,Molecular Biology ,Cells, Cultured - Published
- 1977
194. Monoclonal antibodies to intermediate filaments in chick muscle cell cultures
- Author
-
Linda P. Head and Glenn E. Morris
- Subjects
Hot Temperature ,Fluorescent Antibody Technique ,Muscle Proteins ,Vimentin ,Chick Embryo ,macromolecular substances ,chemistry.chemical_compound ,Intermediate Filament Proteins ,Antibody Specificity ,Animals ,Myocyte ,Isoelectric Point ,Intermediate filament ,Cells, Cultured ,Cytoskeleton ,Immunosorbent Techniques ,Actin ,biology ,Colcemid ,Myogenesis ,Muscles ,Demecolcine ,Antibodies, Monoclonal ,Cell Biology ,Molecular biology ,Molecular Weight ,chemistry ,biology.protein ,Desmin ,Myofibril - Abstract
Two monoclonal antibodies, FIFI and PHIL, have been prepared using detergent-washed myogenic cells as immunogen. On Western blots of total protein extracts of muscle cells, both antibodies bind to vimentin (52 kD) and its degradation products (major band at 42 kD), but do not bind to mouse proteins or to actin (42 kD). Specificity for a determinant common to vimentin and desmin was confirmed by 2-D gel electrophoresis of muscle cell extracts and purified desmin. Western blots with FIFI reveal particularly well the extreme sensitivity of intermediate filaments (IFs) to proteolysis, which was preventable in brain tissue only by boiling in 1% SDS, although it could be reduced in both brain and muscle by less extreme methods. Western blots suggest a large increase in IF content of differentiating myoblast cell cultures at the time of cell fusion and an increase of at least 4-fold is confirmed by a quantitative immunoassay using a direct ELISA method. Immunofluorescence microscopy shows that this increase is due to the appearance of high concentrations of the intermediate filament antigen at the ends of early myotubes, preceding the appearance of cross-striations in myofibrils. Furthermore, whereas the polar filaments detected by FIFI run right to the ends of the early myotubes and only sparingly penetrate the central area, cross-striated myofibrils (as detected by the monoclonal antibody, SAM) run the length of the myotube but do not reach the ends. Colcemid and colchicine cause the vimentin filaments in fibroblasts to collapse into perinuclear rings or caps, but do not have this effect on the polar fluorescence in early myotubes. Heat shock (2 h at 45 °C) has a similar differential effect. The results suggest that early in muscle differentiation intermediate filament proteins accumulate rapidly at myotube ends, where they are organized differently from those in fibroblasts.
- Published
- 1985
195. Protein kinase and cyclic AMP levels in differentiating myoblasts are altered by extracellular calcium
- Author
-
Nguyen Thi Mân and Glenn E. Morris
- Subjects
Biophysics ,Chick Embryo ,Biochemistry ,Ca2+/calmodulin-dependent protein kinase ,Cyclic AMP ,medicine ,Extracellular ,Animals ,Myocyte ,Protein kinase A ,Molecular Biology ,Cell Nucleus ,biology ,Akt/PKB signaling pathway ,Chemistry ,Muscles ,Skeletal muscle ,Cell Differentiation ,Cell Biology ,Kinetics ,medicine.anatomical_structure ,biology.protein ,Calcium ,Creatine kinase ,Protein Kinases ,cGMP-dependent protein kinase - Abstract
Protein kinase specific activities and cyclic AMP levels show a similar pattern of response, when the Ca2+ concentration is altered in the culture medium of differentiating chick skeletal muscle cells; an increase at intermediate Ca2+ concentrations (0.05–0.2mM), followed by a decrease at higher concentrations (2mM). Effects of Ca2+ on protein kinase appear to be on cyclic AMP-independent enzymes in both nucleus and cytoplasm, and are quite the reverse of Ca2+ effects on the muscle-specific enzyme, creatine kinase.
- Published
- 1980
196. RNA synthesis and the stimulation of insulin biosynthesis by glucose
- Author
-
Glenn E. Morris and A. Korner
- Subjects
business.industry ,Insulin ,medicine.medical_treatment ,Biophysics ,Stimulation ,Cell Biology ,Biochemistry ,chemistry.chemical_compound ,Text mining ,Biosynthesis ,chemistry ,Structural Biology ,Genetics ,medicine ,business ,Molecular Biology - Published
- 1970
197. Myosin Messenger RNA: Studies on Its Purification, Properties and Translation during Myogenesis in Culture
- Author
-
Glenn E. Morris, Katherine Tepperman, Arthur W. Rourke, Stuart M. Heywood, Elizabeth A. Buzash, and William C. Thompson
- Subjects
Genetics ,Messenger RNA ,Myogenesis ,Chemistry ,Myosin ,Translation (biology) ,Molecular Biology ,Biochemistry ,Cell biology - Published
- 1973
198. Monoclonal antibody evidence for structural similarities between the central rod regions of actinin and dystrophin
- Author
-
M. M. B. Van Paassen, I. B. Ginjaar, Glenn E. Morris, G.J.B. van Ommen, J.M. Ellis, Antoon F.M. Moorman, Nguyen thi Man, Alison J. Cartwright, and Other departments
- Subjects
Gene isoform ,musculoskeletal diseases ,Monoclonal antibody ,pATH vector ,Blotting, Western ,Biophysics ,Actinin ,macromolecular substances ,Biochemistry ,Dystrophin ,Epitopes ,Mice ,Structural Biology ,Antibody Specificity ,Utrophin ,Genetics ,Animals ,Humans ,Molecular Biology ,Actin ,Antiserum ,biology ,Muscles ,Antibodies, Monoclonal ,Cell Biology ,Muscular Dystrophy, Animal ,musculoskeletal system ,Muscular dystrophy ,Molecular biology ,Molecular Weight ,Actinin, alpha 1 ,Microscopy, Fluorescence ,Polyclonal antibodies ,biology.protein ,Recombinant fusion protein ,Chickens - Abstract
A monoclonal antibody, MANDYS141, binds to both dystrophin and actinin on Western blots (SDS-denatured), but only to actinin in frozen sections of human muscle (native conformation). It differs from a polyclonal cross-reacting antiserum in that it binds to several muscle isoforms of actinin (smooth, fast and slow) from man, mouse and chicken and recognises a quite different part of the proposed triple-helical region of dystrophin (amino acids 1750–2248). The results suggest that structural homologies between actinin and dystrophin occur more than once in their central helical regions and provide experimental support for an actinin-like central rod model for dystrophin.
- Full Text
- View/download PDF
199. Utrophin, the autosomal homologue of dystrophin, is widely-expressed and membrane-associated in cultured cell lines
- Author
-
Nguyen thi Man, LeThiet Thanh, Glenn E. Morris, Derek J. Blake, and Kay E. Davies
- Subjects
Utrophin ,Cell ,Blotting, Western ,Biophysics ,Membrane-associated protein ,Cell Fractionation ,Biochemistry ,Dystrophin-related protein ,HeLa ,Dystrophin ,Mice ,Structural Biology ,Chlorocebus aethiops ,Genetics ,medicine ,Animals ,Humans ,Molecular Biology ,Cells, Cultured ,biology ,Membrane Proteins ,Cell Biology ,biology.organism_classification ,musculoskeletal system ,Muscular dystrophy ,Molecular biology ,Rats ,Blot ,Cytoskeletal Proteins ,medicine.anatomical_structure ,Membrane protein ,Cell culture ,biology.protein ,Cell fractionation ,DMDL gene ,HeLa Cells - Abstract
Utrophin, the autosomal dystrophin-related protein (DRP), is expressed in HeLa cells, smooth muscle-like BC3HI cells from mouse brain, COS monkey kidney cells, the P38BD1 monocyte-macrophage cell line and untransformed human skin fibroblasts, as well as in rat C6 glioma and Schwannoma cells. It was undetectable, however, in the Sp2/O mouse myeloma cell line and in hybridoma lines derived from it. Dystrophin was not detected in any of these cell lines. Although all utrophin-containing cells were capable of forming monolayers in culture, no major effects of either attachment to substratum or length of time in culture (2–17 days) on utrophin levels were observed. After subcellular fractionation ofBC3HI or glioma cells, nearly all of the utrophin was found in the Triton-soluble fraction, suggesting an association with cell membranes.
- Full Text
- View/download PDF
200. Phosphorylation of nuclear proteins during muscle differentiation in vitro
- Author
-
Nguyen Thi Mân, R.J. Cole, and Glenn E. Morris
- Subjects
Time Factors ,Biophysics ,Muscle Proteins ,Chick Embryo ,Biochemistry ,Surface-Active Agents ,Adenosine Triphosphate ,Structural Biology ,Genetics ,Cyclic AMP ,Animals ,Nuclear protein ,Molecular Biology ,Cyclic GMP ,Cells, Cultured ,Cell Nucleus ,Chemistry ,Muscles ,Cell Differentiation ,Cell Biology ,Phosphoproteins ,In vitro ,Cell biology ,Kinetics ,Nucleoproteins ,Isotope Labeling ,Phosphorylation ,Spectrophotometry, Ultraviolet ,Phosphorus Radioisotopes - Full Text
- View/download PDF
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