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Research Interests:
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Publisher Summary The peptidyltransferase of the ribosome has several catalytic activities. One of these is the familiar function in normal peptide bond synthesis. In that process, a peptide group is transferred from one tRNA molecule to... more
Publisher Summary The peptidyltransferase of the ribosome has several catalytic activities. One of these is the familiar function in normal peptide bond synthesis. In that process, a peptide group is transferred from one tRNA molecule to the a-amino group of an adjoining aminoacyl-tRNA, resulting in the formation of an amide or peptide linkage. Several assays have been developed for measuring the ability of the ribosomal peptidyltransferase to form peptide linkages. This chapter describes an assay for the ability of the ribosomal peptidyltransferase to form ester linkages. In experiments, the α-amino group of aminoacyl-tRNA is converted chemically to an α-hydroxy group. In this form, the deaminated aminoacyl-tRNA can be used to form ester linkages and polyester linkages, directed either by synthetic polynucleotides or by naturally occurring messenger RNA. In a closely related reaction, an analog of puromycin is used in which the α-amino group has substituted for it an α-hydroxy group. In that form the α-hydroxy puromycin can be used in a modification of the fragment reaction. In the fragment reaction, formylmethionyl puromycin is formed. When the analog is used, the formylmethionyl group is linked to the puromycin residue by an ester linkage.
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Publisher Summary This chapter discusses the cloning of streptococcal protein G genes. The DNA sequences of the cloned genes have provided the complete amino acid sequences of the proteins they encode, facilitating the dissection of their... more
Publisher Summary This chapter discusses the cloning of streptococcal protein G genes. The DNA sequences of the cloned genes have provided the complete amino acid sequences of the proteins they encode, facilitating the dissection of their structure and function. The comparison of the different genes has elucidated the mechanisms of variation among streptococcal isolates. To facilitate the screening of protein G-producing clones, colony immunoassay procedure is used. The principle of this procedure is that the cellulose acetate filter retains the cells but allows any protein that is released from them to pass through onto the nitrocellulose filter, where it is adsorbed. The adsorbed protein can then be located on the nitrocellulose with high sensitivity by any of a variety of immunochemical staining procedures. The result is an image of the overlying protein-releasing colonies, with very little distortion or loss of detail due to diffusion. The sectoring of colonies is readily apparent when there is instability and subtle differences can be distinguished in the intensity of the staining reaction, reflecting differences in the level of protein release.
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Protein G of Streptococcus is an immunoglobulin-binding protein analogous to protein A of Staphylococcus aureus . Because it binds to several animal IgG's and IgG subclasses (including human IgG3) to which protein A does not bind,... more
Protein G of Streptococcus is an immunoglobulin-binding protein analogous to protein A of Staphylococcus aureus . Because it binds to several animal IgG's and IgG subclasses (including human IgG3) to which protein A does not bind, protein G would be a superior replacement for protein A in many immunochemical applications. To obtain a convenient recombinant source of protein G, genes encoding the protein have been cloned from several independent streptococcal isolates. These cloned genes reveal a structure of protein G which generally resembles protein A, but shows no apparent amino acid sequence homology to protein A in the areas responsible for IgG-binding. The protein is constructed of sets of repeating sequences, the number of which differs in the isolates examined.
Research Interests: Engineering, Technology, Biology, Biological Sciences, HSPA, and 3 moreGene, Recombinant DNA, and Protein A
Deamination of phenylalanyl-transfer RNA with nitrous acid yields the alpha-hydroxyacyl analog, phenyllactyl-transfer RNA. When this is incubated in a protein-synthesizing system directed by polyuridylic acid, it yields an... more
Deamination of phenylalanyl-transfer RNA with nitrous acid yields the alpha-hydroxyacyl analog, phenyllactyl-transfer RNA. When this is incubated in a protein-synthesizing system directed by polyuridylic acid, it yields an acid-precipitable, alkali-labile polyester of phenyllactic acid.
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Research Interests:
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Abstract Functionally active hybrid 50 S ribosomal subunits can be reconstituted using 23 S RNA from Staphylococcus aureus (strain 1206) and 5 S RNA, as well as 50 S ribosomal proteins from Bacillus stearothermophilus. Using this... more
Abstract Functionally active hybrid 50 S ribosomal subunits can be reconstituted using 23 S RNA from Staphylococcus aureus (strain 1206) and 5 S RNA, as well as 50 S ribosomal proteins from Bacillus stearothermophilus. Using this system, resistance of S. aureus 50 S ...
Research Interests: Microbiology, Molecular Biology, Biology, Medicine, Carbon Isotopes, and 14 moreBacillus, Microbial genetic and drug resistance, Staphylococcus aureus, ribosomal RNA, Phenylalanine, Time Factors, Staphylococcus, Erythromycin, Species Specificity, Ribosomes, Protein Binding, Lincomycin, Biochemistry and cell biology, and binding sites
Abstract Two acidic proteins from the 50 S subunit of Bacillus stearothermophilus ribosomes, namely B-L13 (homologous to Escherichia coli protein L 7 L 12 ) and B-L8, form a complex. Radioactive B-L13, added to ribosomes before... more
Abstract Two acidic proteins from the 50 S subunit of Bacillus stearothermophilus ribosomes, namely B-L13 (homologous to Escherichia coli protein L 7 L 12 ) and B-L8, form a complex. Radioactive B-L13, added to ribosomes before dissociation, does not appear in the complex after electrophoresis, so the (B-L13 · B-L8) complex must exist in the ribosome before dissociation. Digestion of B. stearothermophilus ribosomes with polyacrylamide-bound trypsin causes the appearance of new B-L8 and B-L13 spots on two-dimensional polyacrylamide gel electrophoresis, in a pattern which suggests that single molecules of B-L13 are being sequentially cleaved from a four-to-one complex of B-L13 and B-L8.
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Bionanotechnology aims to impart new properties to materials from unique functionalities present in biomolecules. However, the promise of bionanotechnology has not materialized beyond the biomedical field due in large part to issues of... more
Bionanotechnology aims to impart new properties to materials from unique functionalities present in biomolecules. However, the promise of bionanotechnology has not materialized beyond the biomedical field due in large part to issues of scalability, purity, and cost of manufacturing. In this work we demonstrate an approach to co‐engineer production and system functionality into a single polypeptide. We designed a system to anchor particles onto hair via a multifunctional polypeptide composed of two domains, one with affinity to hair and the other capable of strong interactions with the particle surface. These strong interactions, exemplified by resistance to anionic surfactants, stem from the ability to self‐assemble into higher order structures, which were observed by atomic force microscopy. At the same time, the controlled solubility properties of the particle binding domain permit the scalable production in Escherichia coli via inclusion bodies and cost effective purification. We...
Research Interests: Materials Science, Biotechnology, Nanobiotechnology, Industrial Biotechnology, Nanotechnology, and 12 moreAtomic Force Microscopy, Medicine, Environmental Biotechnology, Escherichia coli, Medical Biotechnology, Nanostructures, Peptides, Surface Properties, Biomolecule, Amino Acid Sequence, Silicon Dioxide, and Molecular Sequence Data
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Research Interests: Engineering, Materials Science, Raman Spectroscopy, Electrospinning, Medicine, and 15 moreProtein Engineering, Casting, Genetic Engineering, Biological Sciences, Silk, Animals, CHEMICAL SCIENCES, Spider Silk, Secondary Structure, Protein Secondary Structure Prediction, Fourier transform, Amino Acid Sequence, Biomacromolecules, Molecular Sequence Data, and Fibroins
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Research Interests: Biochemistry, Computer Science, Information Retrieval, Chemistry, Kinetics, and 15 morePhotochemistry, Medicine, Icon, Ribosomal proteins, Citation, Active site, Altmetrics, ICON, Ribosomes, Protein Binding, Ribosomal Protein, Biochemistry and cell biology, binding sites, GTP, and Medical biochemistry and metabolomics
Research Interests: Biochemistry, Computer Science, Chemistry, Crystallography, Medicine, and 15 moreGene expression, Circular Dichroism, Differential scanning calorimetry, Escherichia coli, Bacteria, Citation, Molecular cloning, Altmetrics, G protein, Base Sequence, Binding Site, Biochemistry and cell biology, immunoglobulin G, Molecular Sequence Data, and Medical biochemistry and metabolomics
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The environmental distribution of Dehalococcoides group organisms and their association with chloroethene-contaminated sites were examined. Samples from 24 chloroethene-dechlorinating sites scattered throughout North America and Europe... more
The environmental distribution of Dehalococcoides group organisms and their association with chloroethene-contaminated sites were examined. Samples from 24 chloroethene-dechlorinating sites scattered throughout North America and Europe were tested for the presence of members of the Dehalococcoides group by using a PCR assay developed to detect Dehalococcoides 16S rRNA gene (rDNA) sequences. Sequences identified by sequence analysis as sequences of members of the Dehalococcoides group were detected at 21 sites. Full dechlorination of chloroethenes to ethene occurred at these sites. Dehalococcoides sequences were not detected in samples from three sites at which partial dechlorination of chloroethenes occurred, where dechlorination appeared to stop at 1,2- cis -dichloroethene. Phylogenetic analysis of the 16S rDNA amplicons confirmed that Dehalococcoides sequences formed a unique 16S rDNA group. These 16S rDNA sequences were divided into three subgroups based on specific base substitu...