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Biomedical subjects

J Shine

Publications and source records attributed to J Shine.

154 records · Page 9Linked to original sources
PubMed1974

The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites.

With a stepwise degradation and terminal labeling procedure the 3'-terminal sequence of E. coli 16S ribosomal RNA is shown to be Pyd-A-C-C-U-C-C-U-U-A(OH). It is suggested that this region of the RNA is able to interact with mRNA and that the 3'-terminal U-U-A(OH) is involved in the termination of protein synthesis through base-pairing with terminator codons. The sequence A-C-C-U-C-C could recognize a conserved sequence found in the ribosome binding sites of various coliphage mRNAs; it may thus be involved in the formation of the mRNA.30S subunit complex.

Bacterial Proteins↗
PubMedDate not supplied

Synthesis of growth hormone by bacteria.

A hybrid gene was constructed between the beta-lactamase gene of plasmid pBR322 and the cloned coding sequence for rat growth hormone. This gene is expressed in bacteria and growth hormone sequences are detectable by immunological methods.

Animals↗
PubMedDate not supplied

Structure of a genomic clone encoding biologically active human relaxin.

Relaxin is a peptide hormone synthesized in the corpora lutea of ovaries during pregnancy and is released into the blood stream prior to parturition. Its major biological effect is to remodel the mammalian reproductive tract to facilitate the birth process. Determination of the structure of human relaxin is thus a first step in opening up the possibility of clinical intervention in cases of difficult labour. However, the limited availability of human ovaries during pregnancy has prevented both direct amino acid sequence determination and isolation of cDNA clones obtained from relaxin producing tissue. Our approach has therefore been to screen directly for a human relaxin gene using an homologous porcine relaxin cDNA probe. We report here the successful identification of a genomic clone from which the structure of the entire coding region of a human preprorelaxin gene has been determined. Synthesis of biologically active relaxin has shown that the novel gene structure described herein codes for an authentic human relaxin. We believe this is the first successful synthesis of a biologically active hormone whose structure was predicted solely from the structure of a genomic clone.

Amino Acid Sequence↗
PubMedDate not supplied

Molecular cloning, characterization, and localization of the human homolog to the reported bovine NPY Y3 receptor: lack of NPY binding and activation.

A cDNA clone encoding the human homolog of the bovine cDNA clone LCR1 was isolated from a human lung cDNA library. The 1,670-bp-long nucleotide sequence predicts a single open reading frame of 352 amino acids, with a 92% amino acid identity to a bovine sequence reported to represent the neuropeptide Y (NPY) Y3 receptor. The amino acid sequence shares features common to many other G-protein-coupled receptors, including the seven transmembrane regions and putative glycosylation and phosphorylation sites. Polymerase chain reaction (PCR) analysis of human-hamster hybrid cell DNA reveals that the corresponding gene is located on human chromosome 2. Although the ligand for the bovine receptor has previously been identified as NPY in binding studies, extensive analysis with the human homolog transfected in several different cell lines failed to confirm this classification. Furthermore, the receptor shows 36% identity to both the human interleukin-8 (IL-8) and angiotensin II receptors but only 21% identity to the human NPY Y1 receptor. In addition, NPY and a number of other ligands fail to induce any change in cytosolic calcium levels in transfected cells, suggesting that this clone represents a novel neuropeptide receptor.

Amino Acid Sequence↗
PubMedDate not supplied

Synthesis of mouse renin as a 2-5-33-5 kilodalton pre-pro-two-chain molecule and use of its cDNA to identify the human gene.

A DNA coding for mouse submandibular gland (MSG) renin was used in studies of the biosynthesis and processing of MSG renin and precursors and as a probe for identification of the human renin gene. A 2 kilodalton signal peptide was demonstrated by cleavage of preprorenin (45 kilodaltons) in the presence of microsomal membranes from dog pancreas. Prior selection of renin mRNA by hybridization with its cDNA obviated the possibility that a protease translated from total MSG mRNA could be activated by the microsomal membranes and then act on the primary translation product. In vitro labelling experiments with female MSG demonstrated that prorenin (43 kilodaltons) is rapidly converted to renin (38 kilodaltons) and that testosterone stimulated synthesis by increasing transcription of renin mRNA. Electrophoresis under non-reducing conditions demonstrated 38 kilodalton renin which when reduced ran as two bands of 33 and 5 kilodaltons. Thus, native MSG renin has two chains linked by disulphide bonding. Hydrolysis of the 38 kilodalton single chain occurred only slowly during in vitro labelling. Prorenin bound only weakly to a pepstatin affinity column and could be activated by adding trypsin to column fractions. Both 38 kilodalton single and two chain renin bound strongly however, suggesting that both are active. Base sequencing of MSG renin cDNA indicated an Arg-Arg residue additional to the published amino acid sequence of the heavy chain. This may account for the two forms seen on isoelectric focusing. Mouse renin cDNA was used as a hybridization probe in screening a human genomic library in order to identify the human renin gene. The DNA in positive colonies had overlapping restriction maps and the coding region was found.

Animals↗
PubMedDate not supplied

Structure of human renin and expression of the renin gene.

The amino acid sequence of human renin was identified for the first time. This was determined from the nucleotide sequence of exons in the human renin gene identified in a genomic library by recombinant DNA techniques. Examination of amino acid residues involved in the enzymatic hydrolysis by human renin of the unique Leu10-Val11 bond of human angiotensinogen revealed features peculiar to this highly specialized aspartyl protease. The expression of the renin gene was examined with a hybridization probe for renin mRNA in sections and extracts of tissues. In the submandibular gland of mice renin mRNA, like renin, increased during development and in response to testosterone in females; sodium depletion increased renin mRNA in kidney.

Amino Acid Sequence↗
PubMedDate not supplied

Biomechanics of ankle ligament reconstruction. An in vitro comparison of the Broström repair, Watson-Jones reconstruction, and a new anatomic reconstruction technique.

We wanted to use biomechanical testing in a cadaveric model to compare the Broström repair, the Watson-Jones reconstruction, and a new anatomic reconstruction method. Eight specimens were held in a specially designed testing apparatus in which the ankle position (dorsiflexion-plantar flexion and supination-pronation) could be varied in a controlled manner. Testing was done with intact ligaments and was repeated after sectioning of the anterior talofibular ligament and the calcaneofibular ligament and after a Broström repair, a Watson-Jones reconstruction, and a new anatomic reconstruction were performed. An anterior drawer test was performed using an anterior translating force of 10 to 50 N, and a talar tilt test was performed using a supination torque of 1.1 to 3.4 N-m. The forces in the anterior talofibular ligament and calcaneofibular ligament were measured with buckle transducers, and tibiotalar motion and total ankle joint motion were measured with an instrumented spatial linkage. The increase in ankle joint laxity observed after sectioning of both the anterior talofibular and calcaneofibular ligaments was significantly reduced by the three reconstructive techniques, although not always to the level of the intact ankle. Joint motion was restricted after the Watson-Jones procedure compared with that in the intact ankle. Unlike the Watson-Jones procedure, the ligament or graft force patterns observed during loading after the Broström repair and the new anatomic technique resembled those observed in the intact ankle.

Adult↗